Biomarkers of ionizing radiation

ABSTRACT

The present invention provides novel radiation associated markers. The radiation associated markers may be one or more of albumin, LTGF-β, or any protein or peptide listed in any one of Tables 1, 2, 3, 4, 5, and 6 provided herein. The present invention also provides methods of assessing exposure to ionizing radiation by determining the presence of one or more radiation associated markers. The methods may optionally include quantifying one or more of the radiation associated markers. The methods may further include comparing the amount of one or more radiation associated markers in the sample determined to be present in the sample with either (i) the amount determined for temporally matched, normal samples or (ii) the amount determined for samples obtained from individuals or subjects that have not been exposed to an elevated level of ionizing radiation.

FIELD OF THE INVENTION

This invention relates to new protein biomarkers that may be presentafter exposure to ionizing radiation and methods of assessing exposureto ionizing radiation as well as diagnostic tests and kits forevaluating exposure to ionizing radiation.

BACKGROUND OF THE INVENTION

Recent work has identified a transcription factor, nuclear factor KAPPAB (NF-KB) which induces the TNF-α encoding gene and activates thecyclooxygenase-2 (COX-2) pathway. At 24 hours post irradiation HIF-1aand COX-2 protein levels were increased. In addition to its wellestablished DNA-damage effects, ionizing radiation induces cell death,and radiation-induced activation of acid sphingomyelinases (ASMases) andthe generation of ceramide. Ceramide is generated from sphingomyeline bythe action of a neutral or ASMase or by de novo synthesis coordinatedthrough the enzyme ceramidesynthase. Once generated, ceramide may serveas a second messenger molecule in signaling responses to physiologic orenvironmental stimuli, or it may be converted to a variety of structuralor effector molecules. With a single dose of 3 Gy, there is activationof protein kinase B/AKT (PKB/AKT) signaling. Within minutes ofirradiation, phosphorylation of the serine/threonin protein kinasePKB/AKT at serine-residue 473 appears. This activation of PKB/AKTcontributes to inhibit glycogen synthase kinase-3beta (GSK3beta), whichhas a clear inhibitory role in endothelial cell survival.

One problem first responders face in the event of a nuclear disaster isto rapidly and efficiently identify victims that need medical treatmentand determine what level of treatment is appropriate. It is a priorityto minimize time to diagnosis to identify patient treatment needs,minimize the time required for emergency technicians (ET) providediagnosis, minimize the number of interactions with the patient,minimize system stockpile cost, minimize the dependence oninfrastructure (communications, transport, etc.), minimize the needs fortrained personnel to administer tests, and uniquely identify patient anddiagnostic results.

Simple, quick test strips that change color, like the ones used forsugar or albumin by diabetics, are currently available. They are usedwithout medical or other highly trained personal or expensive equipment.It would be highly desirable to provide such a simple test strip thatchanges color that could be used as evidence of ionizing radiation (IR)exposure. A positive test that could be used to identify a person whohas been exposed to radioactive material has many applications.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides novel radiationassociated markers. The radiation associated markers may be one or moreof albumin, LTGF-β, or any protein or peptide listed in any one ofTables 1, 2, 3, 4, 5, and 6 provided herein.

In a second aspect, the present invention provides methods of assessingexposure to ionizing radiation by determining the presence of one ormore radiation associated markers. The radiation associated markers maybe one or more of albumin, LTGF-β, or any protein or peptide listed inany one of Tables 1, 2, 3, 4, 5, and 6 provided herein.

The sample used for determining the presence of one or more radiationassociated markers may be, for instance, saliva, a buccal swab, amnioticfluid, plasma, serum, urine or blood. The one or more radiationassociated markers may be identified by many methods well known to thoseof skill in the art including Matrix assisted laser desorptionionization (MALDI) mass spectrometry. The radiation associated markersmay also be identified by contacting the sample with an antibody whichspecifically binds to the radiation associated marker under conditionspermitting formation of a complex between the antibody and the radiationassociated marker, and optionally measuring the amount of complexesformed, thereby determining the amount of the radiation associatedmarker.

The methods may optionally include quantifying one or more of theradiation associated markers. The methods may further include comparingthe amount of one or more radiation associated markers in the sampledetermined to be present in the sample with either (i) the amountdetermined for temporally matched, normal samples or (ii) the amountdetermined for samples obtained from individuals or subjects that havenot been exposed to an elevated level of ionizing radiation. Therelative absence of one or more of the radiation associated markers inthe sample indicates that the individual has not been exposed to anelevated level of ionizing radiation, and the relative abundance of oneor more of the radiation associated markers in the sample indicates thatthe individual has been exposed to an elevated level of ionizingradiation. In some embodiments, one or more of the radiation associatedmarkers are not present in detectable levels in normal tissue unaffectedby elevated levels of ionizing radiation at all. In other embodiments,one or more of the radiation associated markers is present in levelsthat are 10%, 20%, 30%, 50%, 75% or more greater than the amount of thecorresponding radiation associated marker in normal tissue unaffected byelevated levels of ionizing radiation at all. In still otherembodiments, one or more of the radiation associated markers is presentin levels that are 2×, 3×, 4×, 5×, 10×, 25×, 50×, 100×, 1,000×, or even10,000× or more greater than the amount of the corresponding radiationassociated marker in normal tissue unaffected by elevated levels ofionizing radiation at all.

In some embodiments, the elevated levels of ionizing radiation may be10%, 20%, 30%, 50%, 75% or more greater than the amount of ionizingradiation experienced normally from environmental sources. In stillother embodiments, the elevated levels of ionizing radiation may be 2×,3×, 4×, 5×, 10×, 25×, 50×, 100×, 500×, 1,000×, 5,000×, 10,000×100,000×,1,000,000× or more greater than the amount of ionizing radiationexperienced normally from environmental sources.

The methods of the present invention are applicable to determining therelative amount of ionizing radiation to which an individual has beenexposed. Likewise, the methods of the present invention are applicableto determining the relative amount of damage caused by ionizingradiation to an individual. The relative amount of ionizing radiation towhich an individual has been exposed and the relative amount of damagecaused by ionizing radiation to an individual may be determined, forinstance, by quantifying the amount of one or more radiation associatedmarker present in a tissue or by determining the presence of one or moreradiation associated marker that is not present in a tissue until aparticular threshold of ionizing radiation is experience or until aparticular threshold of damage to a tissue is crossed.

In a third aspect, the present invention further provides a method ofpredicting outcome in a subject after exposure to elevated levels ofionizing radiation. In some embodiments, it is a method of predictingthe likelihood of surviving long term after exposure to elevatedionizing radiation. In other embodiments, it is a method of predictingthe relative amount of damage caused to an individual or to a tissue byexposure to elevated ionizing radiation comprising determining thepresence or the relative presence as compared to normal samples ornormal subjects of one or more radiation associated markers. One, two,three, or more of the radiation associated markers may be present in thesample and may be identified. One, two, three, or more of the radiationassociated markers may be present in the sample and may be elevated incomparison to samples obtained from individuals that have not beenexposed to elevated levels of ionizing radiation.

The sample used for determining the presence of one or more radiationassociated markers may be, for instance, saliva, plasma, serum, urine orblood. The one or more radiation associated markers may be identified bymany methods well known to those of skill in the art including Matrixassisted laser desorption ionization (MALDI) mass spectrometry. Theradiation associated markers may also be identified by contacting thesample with an antibody which specifically binds to the radiationassociated marker under conditions permitting formation of a complexbetween the antibody and the radiation associated marker, and optionallymeasuring the amount of complexes formed, thereby determining the amountof the radiation associated marker in the sample.

The methods may optionally include quantifying one or more of theradiation associated markers. The methods may further include comparingthe amount of radiation associated markers in the sample determined tobe present in the sample with either (i) the amount determined fortemporally matched, normal samples or (ii) the amount determined forsamples obtained from individuals that have not been exposed to elevatedlevels of ionizing radiation. The relative absence of one or more of theradiation associated markers in the sample indicates that the likelihoodof significant damage, death, illness or medical complications isrelatively low, and the relative abundance of one or more of theradiation associated markers in the sample indicates that the likelihoodof significant damage, death, illness or medical complications isrelatively high.

The invention provides methods for determining the relative likelihoodof significant damage, death, illness or medical complicationscomprising the steps of: (a) obtaining a biological sample from anindividual; (b) measuring an amount of one or more radiation associatedmarkers present in the biological sample; and (c) comparing the amountof one or more radiation associated markers with a predetermined value,whereby the amount of radiation associated markers relative to thepredetermined value indicates the likelihood of significant damage,death, illness or medical complications or the relative likelihood of apositive or negative medical outcome.

The methods of the invention can be used alone or in combination withany known test for determining the relative likelihood of significantdamage, death, illness or medical complications or for determining theprognosis of exposure to ionizing radiation, including, but not limitedto, X-Ray, ultrasound, CAT scan, and other blood marker analysis. Themethods of the invention may be used to screen a biological samplecollected at any time either before or after a first exposure toelevated levels of ionizing radiation has occurred.

In a fourth aspect, the present invention further provides a method ofdetermining the amount of radiation therapy that has been delivered to aparticular tissue. In other embodiments, it is a method of predictingthe relative amount of damage caused to a particular tissue by exposureto elevated ionizing radiation from radiation therapy comprisingdetermining the presence or the relative presence as compared to normalsamples or normal subjects of one or more radiation associated markers.One, two, three, or more of the radiation associated markers may bepresent in the tissue and may be identified. One, two, three, or more ofthe radiation associated markers may be present in the tissue and may beelevated in comparison to tissues that have not received radiationtherapy.

In a fifth aspect, the present invention provides a kit for assessingthe likelihood of significant damage, death, illness or medicalcomplications post exposure to elevated levels of ionizing radiation bydetermining the presence or absence or by quantifying the amount of oneor more radiation associated markers. The kits may contain biomarkeridentification and bioassay systems that address both the problem of avery rapid and inexpensive Point of Care (POC) diagnostic determinationof radiation exposure and of quantitative High Throughput (FIT)diagnostic determination of radiation exposure. The kits and bioassaysystems are useful as a means for making a rapid diagnosis and aquantitative high throughput diagnosis. The kits and bioassay systemsutilize biochemistry that serves as the basis for assay systems based onone or more biomarkers indicative of a dose or ionizing radiationreceived. Immunoassays may be provided to quantitatively measureradiation dose. These immunoassays may be incorporated into commercialdiagnostic systems using both an established strip test technology toprovide a >2 Gy screening diagnosis for quick triage and microfluidicstechnology that provides a low cost high throughput quantitationcapability to indicate dosing level at +/−0.5 Gy to determine the courseof treatment to be received. Examples of such microfluidics devicesinclude a handheld microfluidic reader available from ClarosDiagnostics, Inc.

The kits and bioassays use identified biomarkers. Commercial antibodiesmay be obtained or immunoassays developed to provide a quantitativemeasure of the biomarkers. The commercial antibodies or immunoassays maybe embedded into a test strip using established technology. These teststrips may provide, for instance, a 0-5 or 0-2 Gy quantitative screeningdiagnosis for quick triage. In some embodiments, the test strips maychange color to provide a useful gauge of the amount of elevatedionizing radiation to which a tissue or an individual has been exposedor to gauge the amount of damage a tissue or an individual hasexperienced.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 demonstrates the presence of radiation associated biomarkers postexposure to elevated levels of ionizing radiation.

FIG. 2 demonstrates the presence of radiation associated biomarkers postexposure to elevated levels of ionizing radiation.

FIG. 3 represents a MALDI spectrum, linear mode, control, 15 minutespost 3 Gy, and 30 minute post 3 Gy. Note the protein peaks 63870.32 Daat 15 minutes, and 72449.25 at 30 minutes. These proteins can betentatively identified with bioinformatics in the NCBlnr database. Thetip of the 63870.32 peak is superimposed on the bottom of the 30 minutespectrum.

FIG. 4 represents MALDI, reflectron mode, control, bottom; 15 minutespost 3 Gy, center; and 30 minute post 3 Gy, top. Note 955.87 Da and1184.12 Da peptides that appear at 15 minutes post 3 Gy, and the 939.07Da peptide that appears 30 minute post 3 Gy.

FIG. 5 represents a MALDI image of murine tongue one hour post 1 Gy TBI,longitudinal section. Note peripheral foci of Albumin (Red in image).

FIG. 6 depicts the histopathology of murine tongue one hour post 1 GyTBI, longitudinal section. Note minimal destructive changes in theepithelial cornified spicule layer and edema in the basal region.

FIG. 7 represents a MALDI image. Note the marked increase in peripheralfoci of Albumin compared to the post 1 Gy TBI image (Red in image).

FIG. 8 depicts the histopathology of murine tongue one hour post 2 GyTBI, longitudinal section. Note progressive destructive changes in theepithelial layer with disruption of the cornified spicule layer,increased scattered chromatin debris and edema of the sub-basementmembrane layer.

FIG. 9 represents a MALDI image of murine tongue one hour post 3 Gy TBI,longitudinal section. Note loss of peripheral foci of albumin consistantwith destructive changes in the epithelial layer and increase in centralfoci of Albumin compared to the post 2 Gy image.

FIG. 10 depicts the histopathology of murine tongue one hour post 3 GyTBI, longitudinal section. Note progressive destructive changes andvirtual complete loss of the cornified spicule layer corresponding tothe loss of peripheral albumin foci (peripheral zone) in the MALDIimage, increased scattered chromatin debris, and increased edema of thesub-basement membrane layer.

FIG. 11 represents the LAP monomer, light blue in the image.

FIG. 12 represents the LAP monomer, light blue and red in the image.

FIG. 13 represents the LAP monomer, light blue and red in the image.

FIG. 14 represents the Da Hemoglobin Subunit α, and Parathymosinare onlyfound one hour post 1 Gy IR, red in the image.

FIG. 15 depicts a fatty acid-binding protein adipocyte seen 1 hour post1 and 2 Gy IR, red in the image.

FIG. 16 demonstrates one hour post 2 Gy. Triosphosphate isomerase andsuperoxide dismutase are seen one hour post 2 Gy, but not seen in normaltissue or at one hour post 1 Gy. Red in the image.

FIG. 17 demonstrates hemaglobin α chains and BC011074 NID seen one hourpost 3 Gy.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the following terms mean as follows:

As used herein, “radiation associated marker” means any molecule, suchas a protein, peptide or fragment thereof whose presence, absence oramount in absolute quantity or in quantity relative to other moleculesmay be used as evidence of exposure to elevated levels of ionizingradiation. Stated differently, a radiation associated marker may be anymolecule that may be used as a statistically significant predictor ofexposure to elevated levels of ionizing radiation, or that may used as astatistically significant predictor of the amount of ionizing radiationto which an individual has been exposed, or that may be used as astatistically significant predictor or clinical outcome post exposure toelevated levels of ionizing radiation.

As used herein, a “predetermined value” is a standardized value based ona control. For example, a predetermined value can be based on an amountof radiation associated marker that is present in a biological sampleobtained from an individual.

The term “amount” is used within the context of the analytical methodused to measure the different radiation associated markers and mayreflect a number, a concentration, etc., depending upon the analyticalmethod chosen to measure the radiation associated markers.

The term “biological sample,” as used herein, generally refers to urine,saliva, serum, plasma, tears, or amniotic fluid. Buccal mucosa is anespecially useful “biological sample” for the present invention becauseof its known sensitivity to ionizing radiation and easy accessibilityfor study.

The term “detecting” as used herein refers to identifying the presenceof, identifying the presence of in relative amounts relative to anothermolecule or radiation associated marker relative to a predeterminedvalue, or quantifying in absolute amounts.

The term “assessing exposure to ionizing radiation” includes quantifyingthe amount of ionizing radiation to which an individual has beenexposed, quantifying the length of time to which an individual has beenexposed to ionizing radiation, estimating or quantifying the amount ofdamage to a tissue or to an individual after exposure to ionizingradiation, and providing a prognosis of likely clinical sequelae orsurvival probability.

The term “elevated levels of ionizing radiation” encompasses instanceswhere the ionizing radiation is present in amounts that are 10%, 20%,30%, 50%, 75% or more greater than the amount of ionizing radiationexperienced normally from environmental sources. The term “elevatedlevels of ionizing radiation” also encompasses instances where theionizing radiation is present in amounts that are 2×, 3×, 4×, 5×, 10×,25×, 50×, 100×, 500×, 1,000×, 5,000×, 10,000×100,000×, 1,000,000× ormore greater than the amount of ionizing radiation experienced normallyfrom environmental sources. Ionizing radiation may be quantified andexpressed in terms of a gray (Gy). One gray is the absorption of onejoule of energy, in the form of ionizing radiation, by one kilogram ofmatter. For X-rays and gamma rays, these are the same units as thesievert (Sv). To avoid any risk of confusion between the absorbed doseand the equivalent dose, one must use the corresponding special units,namely the gray instead of the joule per kilogram for absorbed dose andthe sievert instead of the joule per kilogram for the dose equivalent.The gray measures the deposited energy of radiation. The biologicaleffects vary by the type and energy of the radiation and the organismand tissues involved. The sievert attempts to account for thesevariations. A whole-body exposure to 5 or more grays of high-energyradiation at one time usually leads to death within 14 days. This dosagerepresents 375 joules for a 75 kg adult (equivalent to the chemicalenergy in 20 mg of sugar). Since grays are such large amounts ofradiation, medical use of radiation is typically measured in milligrays(mGy). The average radiation dose from an abdominal x-ray is 1.4 mGy,that from an abdominal CT scan is 8.0 mGy, that from a pelvic CT scan is25 mGy, and that from a selective spiral CT scan of the abdomen and thepelvis is 30 mGy. One gray is equivalent to 100 rad. Therefore, the term“elevated levels of ionizing radiation” may be defined in some instancesto mean a radiation dose greater than 0.25, 0.50, 1.0, 1.5, 2.0, 5.0,10.0, 20.0, 50.0 or 100 mGy. In other instances, the term “elevatedlevels of ionizing radiation” may be defined to mean a radiation dosegreater than 0.25, 0.50, 1.0, 1.5, 2.0, or 5.0 Gy.

According to an embodiment of this invention, the sample may be a salivasample or a buccal swab. The sample may be obtained, for instance, about15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2days or a week after exposure to elevated levels of ionizing radiation.In a particular embodiment of this invention, the sample is a buccalswab.

In addition, the present invention provides a method for determining theamount of radiation associated marker of in a sample comprising: (a)contacting the sample with an antibody which specifically binds to aradiation associated marker under conditions permitting formation of acomplex between the antibody and the radiation associated marker; and(b) determining the amount of complexes formed thereby determining theamount of radiation associated marker in the sample.

This invention is illustrated in the experimental details section whichfollows. These sections are set forth to aid in an understanding of theinvention but are not intended to, and should not be construed to limitin any way the invention as set forth in the claims which followthereafter.

Radiation associated markers can be directly measured, for example,using anti-marker antibodies in an immunoassay, such as a Western blotor ELISA. Radiation associated markers can be indirectly measured, forexample, using a capture antibody that binds the radiation associatedmarker.

The amount of radiation associated markers in a biological sample can bedetermined using any method known in the art, including, but not limitedto, immunoassays using antibodies specific for the radiation associatedmarker. Any assay that functions to qualitatively or quantitativelydetermine variations in sample concentrations of radiation associatedmarkers from normal levels can be employed in the practice of theinvention.

For example, a monoclonal anti-radiation associated marker antibody canbe generated by immunizing a mouse with the radiation associated marker.Once an immune response is detected, e.g., antibodies specific for theradiation associated marker are detected in the mouse serum, the mousespleen is harvested and splenocytes are isolated. The splenocytes arethen fused by well-known techniques to any suitable myeloma cells, forexample, cells from cell line SP20 available from the American TypeCulture Collection (ATCC). Hybridomas are selected and cloned by limiteddilution. The hybridoma clones are then assayed by methods known in theart for cells that secrete antibodies capable of binding the radiationassociated marker. Ascites fluid, which generally contains high levelsof antibodies, can be generated by immunizing mice with positivehybridoma clones.

Any type of fusion phage, monoclonal, or polyclonal antibodies can beused in immunoassays of the invention, so long as the antibodies can beused in a reproducible fashion as markers for various radiationassociated markers or as measures of the different levels of radiationassociated markers observed in normal and variant populations.

In one embodiment, an amount of radiation associated marker can bemeasured using a capture antibody followed by a labeled secondaryantibody using a strategy as described, for example, in U.S. Pat. No.6,429,018, hereby incorporated by reference. The label on the secondaryantibody can comprise any chemical, radioactive, lanthanide, coloreddye, or genetic tag used in enzyme-linked immunosorbent assays (ELISAs),Western blots, and other sensitive and specific immunoassays andimmunoradiometric assays using known methodology. These includeconjugating the antibody with horseradish peroxidase or alkalinephosphatase that are easily measurable, typically using colorimetric,fluorometric or luminescent substrates. Genetic labels include fireflyluciferase, employed because luciferase produces a bioluminescentmolecule when incubated with its substrate, luciferin.

Serum albumin, by test strip, and numerous other novel protein andpeptide biomarkers of ionizing radiation (IR) are consistently found bymass spectrometry in ex vivo wiping and histology sections of murinetongue epithelium one hour post 1, 2, and 3 Gy. Albumin and otherproteins are not seen in non-irradiated tissue. Keratin is a ubiquitouscomponent of tongue tissue, and can be seen at all levels of IR. Theadditional proteins in nonirradiated tissue, 1, 2, and 3 Gy are shown inthe LCMS mass spectrometry Mascot search engine results. Thesebiomarkers can be identified at POC with a simple oral cavity swab.FIG. 1. These biomarkers can be confirmed hours to days after the eventwith the same mass spectrometry techniques used in the initial biomarkeridentification with similar samples.

Matrix-Assisted Laser Desorption/Ionization (MALDI).

Matrix-assisted laser desorption/ionization (MALDI) is a soft ionizationtechnique used in mass spectrometry, allowing the analysis ofbiomolecules (biopolymers such as proteins, peptides and sugars) andlarge organic molecules (such as polymers, dendrimers and othermacromolecules), which tend to be fragile and fragment when ionized bymore conventional ionization methods. It is most similar in character toelectrospray ionization both in relative softness and the ions produced(although it causes many fewer multiply charged ions). The ionization istriggered by a laser beam (normally a nitrogen laser). A matrix is usedto protect the biomolecule from being destroyed by direct laser beam andto facilitate vaporization and ionization.

The matrix consists of crystallized molecules, of which the three mostcommonly used are 3,5-dimethoxy-4-hydroxycinnamic acid (sinapinic acid),α-cyano-4-hydroxycinnamic acid (alpha-cyano or alpha-matrix) and2,5-dihydroxybenzoic acid (DHB). A solution of one of these molecules ismade, often in a mixture of highly purified water and an organic solvent(normally acetonitrile (ACN) or ethanol). Trifluoroacetic acid (TFA) mayalso be added. A good example of a matrix-solution would be 20 mg/mLsinapinic acid in ACN:water:TFA (50:50:0.1). The identity of suitablematrix compounds is determined to some extent by trial and error, butthey are based on some specific molecular design considerations: theyhave a low molecular weight; they are acidic and act as a proton sourceto encourage ionization of the analyte; they have a strong opticalabsorption in the UV and efficiently absorb the laser irradiation; andthey are functionalized with polar groups allowing use in aqueoussolutions.

The matrix solution is mixed with the analyte (e.g. protein sample). Theorganic solvent allows hydrophobic molecules to dissolve into thesolution, while the water allows for water-soluble (hydrophilic)molecules to do the same. This solution is spotted onto a MALDI plate(usually a metal plate designed for this purpose). The solventsvaporize, leaving only the recrystallized matrix, but now with analytemolecules spread throughout the crystals. The matrix and the analyte aresaid to be co-crystallized in a MALDI spot.

The laser is fired at the crystals in the MALDI spot. The matrix absorbsthe laser energy, and the matrix is ionized by this event. The matrixtransfers part of its charge to the analyte molecules (e.g. protein)thereby ionizing them while still protecting them from the disruptiveenergy of the laser. Ions observed after this process consist of aneutral molecule [M] and an added or removed ion. Together, they form aquasimolecular ion, for example [M+H]⁺ in the case of an added proton,[M+Na]⁺ in the case of an added sodium ion, or [M−H]⁻ in the case of aremoved proton. MALDI is capable of creating singly-charged ions, butmultiply charged ions ([M+nH]^(n+)) can also be created, as a functionof the matrix, the laser intensity and/or the voltage used. Note thatthese are all even-electron species. Ion signals of radical cations canbe observed eg. in case of matrix molecules and other stable molecules.

Atmospheric pressure (AP) matrix-assisted laser desorption/ionization(MALDI) is an ionization technique (ion source) that in contrast tovacuum MALDI operates at normal atmospheric environment. In vacuumMALDI, ions are typically produced at 10 mTorr or less while in AP-MALDIions are formed in atmospheric pressure.

AP-MALDI is used in mass spectrometry (MS) in a variety of applicationsincluding proteomics and drug discovery fields. AP-MALDI massspectrometry is ofen used in proteomics, DNA/RNA/PNA, lipids,oligosaccharides, phosphopeptides, bacteria, small molecules andsynthetic polymers, similar applications as available also for vacuumMALDI instruments.

The AP-MALDI ion source is easily coupled to an ion trap massspectrometer or any other MS system equipped with ESI (electrosprayionization) or nanoESI source.

The type of a mass spectrometer most widely used with MALDI is the TOF(time-of-flight mass spectrometer) because of its large mass range. TheTOF measurement procedure is suited to the MALDI ionization processsince the pulsed laser takes individual ‘shots’ rather than working incontinuous operation. MALDI-TOF instruments are typically equipped withan “ion mirror,” deflecting ions with an electric field thereby doublingthe ion flight path and increasing the resolution. Today, commercialreflectron TOF instruments reach a resolving power m/Δm of well above20′000 FWHM (full-width half-maximum, Δm defined as the peak width at50% of peak height).

In proteomics, MALDI is used for identifying proteins isolated throughgel electrophoresis: SDS-PAGE, size exclusion chromatography, andtwo-dimensional gel electrophoresis. One method used is peptide massfingerprinting by MALDI-MS, or with post ionization decay orcollision-induced dissociation.

IMS.

The history of IMS (Imaging Maldi Spectroscopy) began with single cellstudies of Aplysia californica neurons by matrix-assisted laserdesorption/ionization time-of flight mass spectrometry (MALDI/TOF-MS).This was probably the first direct tissue MALDI identification ofpeptides and tissue profiling based on ion density. (Garden et al., JMass Spectrom 1996; 31:1126-30) A refinement of this technique, imagingMALDI was described in human buccal mucosa, and rat pituitary andpancreas glands using two different approaches: direct targeting of thetissue itself and by analysis of blotted targets previously exposed tothe tissue. (Caprioli et al., Anal Chem 1997; 69:4751-60) An “MS ImageTool,” using the peptide neurotensin (peak at m/z 1674) significantlyimproved the data acquisition, speed of IMS, and utilization of thetechnique. (Stoeckli et al., J Am Soc Mass Spectrom 1999; 10:67-71) In alaser, capture microdissection (LCM) study of both tumor and normalhuman breast tissue fixed in ethyl alcohol, and stained with hematoxylinand eosin, normal tissue, carcinoma in situ, invasive carcinoma, andmetastatic carcinoma could be distinguished by their different MALDIspectra. The introduction of “BioMap” software, by applying a baselinecorrection to the spectra and integrating over the peak of interest,demonstrated mouse brain images of amyloid β, and Aβ peptides. Thatreport was proof of the principle that MALDI images of tissue could beobtained based upon the mass spectrometry mass/charge, m/z peak ofinterest. (Stoeckli et al., Nat Med 2001; 7:493-6) Subsequent profilingand IMS of normal mouse epididymis identified different protein activity(ion densities) throughout the sections. (Chaurand et al.,Electrophoresis 2002; 23:3125-35)

The negative effects on IMS resolution from destructive tissue freezingartifacts, excessive dehydration due to ethanol fixation,paraformaldeyde cationization, embedding artifacts from OCT polymer andagar, and coarse matrix crystal size were first described in a report onspatial profiling of invertebrate ganglia. (Kruse et al., J Am Soc MassSpectrom 2003; 14:752-9) In a follow-up report, they suggested asolution to the problem of freezing artifacts using glycerol and therelated compounds ethane-1,2-diol and propane-1,2-diol to stabilizecellular membranes. (Rubakhin et al., Anal Chem 2003; 75:5374-80) Asubsequent report suggested direct liquid nitrogen immersion of tissuein aluminum wrapping as a means of rapid fixation, but ignored the knownconsequences of freezer artifact. (Schwartz et al., J Mass Spectrom2003; 38:699-708) Adjunctive histologic staining with methylene bluestained tissues on standard metal plates or indium-tin coated glassslides were shown to be compatible with IMS, but cresyl violet staindecreased IMS signal intensity. (Chaurand et al., Anal Chem 2004;76:1145-55)

Tissue blotting with trypsin digestion for MSMS (mass spectrometry massspectrometry using two mass spectrometers in tandem) data base analysiswas shown to be useful in analyte localization, but was destructive totissue morphology. (Bunch et al., Rapid Communications in MassSpectrometry 2004; 18:3051-60) Others described a less destructivetrypsin digest step to their prior tissue blotting technique for IMS.(Rohner et al., Mech Ageing Dev 2005; 126:177-85) Another method,matrix-enhanced secondary ion mass spectrometry (ME) SIMS was describedand used for direct molecular imaging of the ganglia of the freshwatersnail, Lymnaea stagnalis. (Altelaar et al., Anal Chem 2005; 77:735-41)However, this technique presented significant limitations and was provedunsuitable for direct tissue imaging. A refinement of IMS, oversamplingwith complete sample ablation at each sample position on the targetplate, provided significant resolution enhancement with a translationstage raster step size of 25 μM. A 40 μM object could now be resolvedwith a 100 μM laser. (Jurchen et al., Journal of the American Societyfor Mass Spectrometry 2005; 16:1654-9)

Tissue treatments with organic solvents such as chloroform, acetone,hexane, toluene, or xylene were shown to be an effective and rapidmethod for signal enhancement in MALDI direct tissue profiling. (Lemaireet al., Analytical Chemistry 2006; 78:7145-53) These studiesdemonstrated that solvent treatments partially removed lipids from thetissue surface. Compared to previous studies with ethanol,chloroform/xylene solvent, rinsing is more specific for lipid removaland does not generate delocalization or extraction of most solublepeptides/proteins as tested by immuno-histochemistry experiments. Amongall the tested solvents, chloroform and xylene produced the greatestincrease in MALDI signal intensity and number of detectedpeptides/proteins. However, this treatment does not reduce salt adductsas does alcohol treatment. The results suggest that it is possible todetect, after organic rinsing treatments, compounds, such aspeptides/proteins present in the cytoplasm, that were masked by lipidsin the tissue.

Clench et al. reported the development and application of a method using9-aminoacridine as a matrix for negatively charged ions in MALDIimaging. (Burrell et al., J Exp Bot 2007; 58:757-63) Crossmandemonstrated the need for thin sections to avoid differential extractionefficiency of matrix solvent in different tissues. (Crossman et al.,Rapid Communications in Mass Spectrometry 2006; 20:284-90) Pevsnerdemonstrated direct cellular MALDI identification of proteins in fixedcells and tissues without freezer artifact, tissue corrosion by matrixsolvents or the use of tissue blotting. This was confirmed in a laterreport. (Pevsner et al., Direct identification of proteins from cellsand tissues using MALDI TOF. Anal Chem.; Pevsner et al., J Soc GynecolInvestig 2006; 13:A1-B10; Groseclose et al., J Mass Spectrom 2007;42:254-62)

Metal-assisted (MetA) secondary ion mass spectrometry (SIMS), avariation on SIMS as well as matrix-assisted laser desorption/ionization(MALDI) IMS can provide images from tissue, but the duration of theseprotocols were highly dependent on sample size and technique parameters.The duration of these studies averaged approximately 5 h. (Altelaar etal., Nat Protoc 2007; 2:1185-96)

Agar et al. studied multiple solvent/matrix combinations. However, thetissue sections at the electron microscopic level demonstrated bothfreezing artifact and structural distortion, indicating that theirmethod disrupts normal subcellular structures such as mitochondria.(Agar et al., Matrix Solution Fixation: Histology-Compatible TissuePreparation for MALDI Mass Spectrometry Imaging. Anal Chem 2007.) Baluyaet al reported a variation on inkjet-printed matrix application totissue specimens previously described by Sloane et al. (Baluya et al.,Anal Chem 2007; 79:6862-7; Sloane et al., Mol Cell Proteomics 2002;1:490-9) The matrix application was of better quality and morereproducible than from specimens prepared by the electrospray andairbrush methods, but still was not completely uniform. A uniform methodof tissue matrix application is sublimation. Sublimation is solventfree, rapid, and was successfully used to identify lipids in braintissue, and more recently described for proteins or peptides. Thechallenge of IMS in formalin fixed paraffin embedded tissue was firstaddressed by and then by Pevsner and later by Stauber. (Hankin et al., JAm Soc Mass Spectrom 2007; 18:1646-52; Pevsner et al. “MicrotubuleAssociated Proteins (MAP) and Motor Molecules: Direct Tissue MALDIIdentification and Imaging.” 2007; Pevsner et al., “Colon Cancer:Protein Biomarkers in Tissue and Body.” 2007; Pevsner et al.,“Colorectal Carcinoma—Field Defects in SatelliteTissue.” 2007;Puolitaival et al., J Am Soc Mass Spectrom 2008; 19:882-6; Lemaire etal., J Proteome Res 2007; 6:1295-305; 48. Stauber et al., J Proteome Res2008; 7:969-78)

This invention is illustrated in the experimental section that follows.These examples are set forth to aid in understanding the invention butare not intended to, and should not be construed to, limit in any waythe invention as set forth in the claims.

Example 1

New protein biomarkers were identified in murine buccal mucosa afterionizing radiation. The new protein biomarkers were identified fromscraping of buccal mucosa obtained fifteen and thirty minutes postionizing radiation exposure. The biomarkers are as follows:

-   -   DnaJ homolog subfamily C member 1 (DnaJ protein homolog MTJ1)        gi|2494160|sp|Q61712.1|DNJC1_MOUSE[2494160. DnaJ domain. DnaJ        domains (J-domains) are associated with hsp70 heat-shock system        and it is thought that this domain mediates the interaction        which is triggered by ionizing radiation.    -   mCG1463, isoform CRA_b [Mus musculus]    -   gi|148683958|gb|EDL15905.1|[14868395]. This protein is involved        in transcriptional regulation which is triggered by ionizing        radiation.        The following three proteins all represent biomarkers of acute        radiation exposure which triggers either transcription or        post-translational modification.    -   Novel protein (B230312A22Rik) [Mus musculus] 955.87 Da    -   Amino Acid Sequence: MSGPRKAPED    -   gi:45593111 Demonstrated 15 minutes post ionizing radiation.    -   Unnamed protein product [Mus musculus] 1184.12 Da    -   Amino Acid Sequence: AKDYYGTGYF    -   gi|1333908| Demonstrated 15 minutes post ionizing radiation.    -   Transaldolase [Mus musculus] 939.07 Da    -   Amino Acid Sequence: MRHVQAELS    -   gi|168984609| Demonstrated 30 minutes post ionizing radiation.

The identification of specific protein biomarker profiles and newprotein biomarkers of ionizing radiation exposure enable reliableassociations with dose-exposure, for biodosimetry purposes. Thisstrategy will result in the development of diagnostic tests using abuccal mucosa swab to provide acute clinical quantitative evidence ofradiation exposure of 3 Gy or more and allow for accurate and immediatetriage of victims of an ionizing radiation mass casualty.

Example 2

Fourteen swiss mice were anesthetized with intraperitonealketamine-xylazine (k/x, 80/10 mg/kg ip). Ten mice received 3 Gy to thehead. Buccal mucosa scrapings were obtained fifteen minutes (5 mice) andthirty minutes (five mice) post exposure. Buccal mucosa scrapings fromthe four non-radiated mice were used as controls. All reagents wereobtained from Sigma (St. Louis, Mo.) and used unmodified. All sampleswere placed in 1 mL of 100 mMolar ammonium bicarbonate buffer. Proteinswere extracted from all the tissues in this buffer with high pressure(Barocycler, Pressure BioSciences, South Easton, Mass.). The sampleswere concentrated by lypholization to 200 μL and divided into threecomponents. One for trypsin digestion and liquid chromatography massspectrometry, LCMS, analysis (bottoms-up proteomics), one for HPLCprotein separation and FTMS analysis (top-down proteomics) to identifypost-translational modifications (this aliquot was frozen for lateranalysis), and one for direct identification with matrix assisted laserdesorption mass spectrometry, MALDI analysis. The study was conductedunder an approved protocol of the New York University School of MedicineInstitutional Animal Care and Use Committee, Laboratory Animal Protocol#080601-01.

Sinapic acid (5 mg/mL) 0.3 μL was pipetted onto a conductive MALDIplate, allowed to dry, and covered with an equal amount of sample. Allfourteen samples were examined with MALDI in linear mode. The findingswere consistent across all four controls, 15 minute (N=5), and 30 minute(N=5) samples, FIG. 3. Cyano 4 hydroxy cinnamic acid (5 mg/mL) 0.3 μLwas pipetted onto a conductive MALDI plate, allowed to dry, and coveredwith an equal amount of sample. All fourteen samples were examined withMALDI in reflectron mode. The findings were consistent across all fourcontrols, 15 minute (N=5), and 30 minute (N=5) samples, FIG. 4.

Results.

The linear mode MALDI MS experiment is shown in FIG. 3. The lowestspectrum is a control sample. The middle spectrum is a 15 minute post 3Gy sample. The upper spectrum is a 30 minute post 3 Gy sample. At 15minutes post 3 Gy, a protein 63870.32 Da, D naJ homolog subfamily Cmember 1 (DnaJ protein homolog MTJ1)gi|2494160|sp|Q61712.1|DNJC1|_MOUSE[2494160] was demonstrated. DnaJdomains (J-domains) are associated with hsp70 heat-shock system and itis thought that this domain mediates the interaction. At 30 minutes post3 Gy, a protein 72449.25 Da, mCG 1463, isoform CRA_b [Mus musculus]gi|148683958|gb|EDL15905.1|[14868395] is tentatively identified in thespectrum. This protein is involved in transcriptional regulation. At 15and 30 minutes post 3 Gy, the spectrum of peptides is demonstrated inFIG. 4, and the peptide amino acid sequences derived with bioinformaticsand the corresponding proteins they represent are shown in Table 1.

Table 1.

Peptide amino acid sequences and corresponding proteins identified withbioinformatics at 15 and 30 minutes post 3 Gy.

These experiments demonstrated novel changes 15 and 30 minutes postionizing radiation to the head and neck.

Example 3

Additional experiments using total body ionizing radiation (N=21), TBI,exposure at 0 (N=3), 1 (N=6), 2 (N=6), and 3 (N=6) Gy were performed(mice). The murine tongue was chosen for examination. The mice weresacrificed at one hour post TBI. MALDI imaging of tongue tissue wascombined with high resolution, 1.43 nm, (IR filter visible light 400 nm,Zeiss 63× oil immersion 1.4 NA objective, and 1.4 NA Olympus condenser)bright field microscopy (the smallest mitochondria are 1000 nm); andliquid chromatography mass spectrometry, LCMS, to obtain proteinsequence data for exact identification and localization of post TBIprotein biomarkers in the tissue sections. Protein extraction from the3^(rd) contiguous section was performed with a Barocycler and digestedwith trypsin for LCMS examination. Contiguous 5μ cryo-sections wereobtained for MALDI imaging, examples of normal, post 1, 2, and 3 Gy,FIGS. 5-10; histology (hematoxylin and eosin, H&E), examples of normal,post 1, 2, and 3 Gy, FIGS. 5-10; and nanoflow liquid chromatography massspectrometry, LCMS, examples of normal, post 1, 2, and 3 Gy. Themolecular weights of the identified proteins were used in the MALDIimage prepared with sublimation for tissue localization. (Naftolin etal., Reproductive Sciences 2007; 14:257A; Pevsner et al., Biomarkers Med2009; 3:55. FIG. 5 represents a MALDI image of murine tongue one hourpost 1 Gy TBI, longitudinal section. Note peripheral foci of Albumin(Red in image). FIG. 6 demonstrates the histopathology of murine tongueone hour post 1 Gy TBI, longitudinal section. Note minimal destructivechanges in the epithelial cornified spicule layer and edema in the basalregion. FIG. 7 represents a MALDI image Note marked increase inperipheral foci of Albumin compared to the post 1 Gy TBI image (Red inimage).

FIG. 8 demonstrates the histopathology of murine tongue one hour post 2Gy TBI, longitudinal section. Note progressive destructive changes inthe epithelial layer with disruption of the cornified spicule layer,increased scattered chromatin debris and edema of the sub-basementmembrane layer. FIG. 9 represents a MALDI image of murine tongue onehour post 3 Gy TBI, longitudinal section. Note loss of peripheral fociof albumin consistant with destructive changes in the epithelial layerand increase in central foci of Albumin compared to the post 2 Gy image.FIG. 10 demonstrates the histopathology of murine tongue one hour post 3Gy TBI, longitudinal section. Note progressive destructive changes andvirtual complete loss of the cornified spicule layer corresponding tothe loss of peripheral albumin foci (peripheral zone) in the MALDIimage, increased scattered chromatin debris, and increased edema of thesub-basement membrane layer.

Table 2.

Table 2 provides the results of a Mascot search of proteins in normalmurine tongue. No albumin was identified.

Table 3.

Table 3 lists LCMS identified proteins with Mascot search of proteins inmurine tongue one hour post 1 Gy TBI. Albumin appears at the end of thelist and corresponds to the findings in the MALDI image one hour post 1Gy.

Table 4.

Table 4 lists LCMS identified proteins with Mascot search of proteins inmurine tongue one hour post 2 Gy TBI. Albumin appears next to last inthe list.

Table 5.

Table 5 lists LCMS identified proteins from two separate samples 16 and19. Mascot search of proteins in murine tongue one hour post 3 Gy TBI.The absence of albumin in these two 3 Gy samples, it was present inother 3 Gy samples (data not shown) is consistent with the marked tissuedestruction, and very small amount noted in the MALDI image.

Example 4

It has been shown (MHB) that latent transforming growth factor-β,LTGF-β, is rapidly activated in vivo after ionizing radiation (IR). Thisis evidenced by increased immunoreactivity of TGFβ epitopes that aremasked by the latent complex. (Barcellos-Hoff Mol Biol Cell 1994;5:139a; Barcellos-Hoff et al., Am J Pathol 1995; 147:1228-37;Barcellos-Hoff et al., Molec Endocrin 1996; 10:1077-83; Barcellos-Hoffet al., Mol Endocrinol 1996; 10:1077-83; Barcellos-Hoff J Mammary GlandBiol Neoplasia 1996; 1:353-63; Ehrhart et al., FASEB J 1997;11:991-1002; Barcellos-Hoff et al., Breast Cancer Res 2000; 2:92-9;Barcellos-Hoff et al., Nat Rev Cancer 2005; 5:867-75; Jobling et al.,Radiat Res 2006; 166:839-48; Jobling et al., Radiat Res 2006;166:839-848). Because reactive oxygen species, ROS, are a product of theinteraction of IR with water or with cell membranes, it was postulatedthat the rapid activation of TGFβ in vivo could be due to ROS generatedby IR if the protein itself contained redox sensitive amino acids. Wewere able to demonstrate the monomers of TGFβ and the latency associatedprotein, LAP, in MALDI images one hour post 1, 2, and 3 Gy, FIGS. 18,19, 20. Note the approximately 10 Da variance in all three images. Thismolecular weight variance and spatial resolution can be improved, infravida. Therefore TGFβ can also be used as a protein biomarker of absorbeddose. Quantization can be obtained from trypsin digests studied by LCMS.FIGS. 11, 12 and 13 depict LAP monomers, light blue in the image.

Additional dose specific protein biomarkers identified from other LCMSstudies were not seen in normal tissue. Hemoglobin subunit α, andparathymosinare only found one hour post 1 Gy; Fatty Acid BindingProtein Adipocyte seen 1 hour post 1 and 2 Gy IR; and triosphosphateIsomerase, and superoxide dismutase seen at one hour post 2 Gy are notseen in normal tissue or at one hour post 1 Gy.

FIG. 14 depicts da hemoglobin subunit a, and parathymosinare only foundone hour post 1 Gy IR, red in the image. FIG. 15 depicts fattyacid-binding protein adipocyte seen 1 hour post 1 and 2 Gy IR, red inthe image. FIG. 16 depicts one hour post 2 Gy exposure. Triosphosphateisomerase and superoxide dismutase are seen one hour post 2 Gy, but notseen in normal tissue or at one hour post 1 Gy (red in the image). FIG.17 demonstrates hemaglobin a chains and BC011074 NID seen one hour post3 Gy exposure.

The results with MALDI imaging, high resolution microscopy, and LCMS areproof of principle that post TBI protein biomarkers can be identifiedand used as measures of absorbed radiation dose at 1, 2, and 3 Gy withour current technology. Identification of a larger library of post TBIproteins will improve specificity of both the POC triage kit and the HTdevice. This will dramatically limit false positives and falsenegatives. This will be achieved with a combination of strategiesincluding higher resolution bright field microscopy, atomic forcemicroscopy of irradiated tissue for higher spatial resolution of tissueloci of proteins by co-registration with the MALDESI images. (Collier etal., Anal Chem 2008; 80:4994-5001; Bereman et al., Rapid Commun MassSpectrom 2008; 22:1563-6; Sampson et al., J Am Soc Mass Spectrom 2009;20:667-73). Separation of extracted proteins is performed by 2D cationexchange LCMS with high-throughput electron capture dissociation (ECD)analysis to identify post translational modification of IR inducedprotein biomarkers; modifying sublimation matrix application by varyingtime, temperature and distance between the matrix powder platform andthe mounted tissue, FIG. 16; thinner tissue sections with a highperformance cryostat; and complete removal of tissue embedding mediapolymer residue by varying solvent baths.

The application of matrix assisted laser distortion electro-sprayionization Fourier transform ion cyclotron mass spectrometry, FTICR, forboth liquid samples and imaging, MALDESI imaging, will improve theisotopic resolution by 2 orders of magnitude. The MALDI system fullwidth half mass, FWHM, isotopic resolution is 2500 Da. The MALDESIsystem full width half mass, FWHM, isotopic resolution is 250,000 Da.The spatial resolution of MALDI images will improve with laser M² beamquality closer to 1, increased repetition rate, Hz, fluence, mj/s(Watts), Energy, E, mj/pulse, pulse width, increased work function,smaller spot size, and, optics (beam focus); and decremental samplestage movement from microns to nanometers.

The following Table 6 provides examples of other potential biomarkers ofionizing radiation isolated from buccal tissue in subsequent examplesobtained according to the procedures set forth in Example 3.

TABLE 6 K1C10_MOUSE Keratin, type I cytoskeletal 10 OS = Mus musculus GN= Krt10 PE = 1 SV = 3 ALBU_MOUSE Serum albumin OS = Mus musculus GN =Alb PE = 1 SV = 3 K2C75_MOUSE Keratin, type II cytoskeletal 75 OS = Musmusculus GN = Krt75 PE = 1 SV = 1 K2C1_MOUSE Keratin, type IIcytoskeletal 1 OS = Mus musculus GN = Krt1 PE = 1 SV = 4 K1C14_MOUSEKeratin, type I cytoskeletal 14 OS = Mus musculus GN = Krt14 PE = 1 SV =2 K2C5_MOUSE Keratin, type II cytoskeletal 5 OS = Mus musculus GN = Krt5PE = 2 SV = 1 K1C15_MOUSE Keratin, type I cytoskeletal 15 OS = Musmusculus GN = Krt15 PE = 1 SV = 2 K1C13_MOUSE Keratin, type Icytoskeletal 13 OS = Mus musculus GN = Krt13 PE = 2 SV = 2 K2C6A_MOUSEKeratin, type II cytoskeletal 6A OS = Mus musculus GN = Krt6a PE = 2 SV= 3 K2C4_MOUSE Keratin, type II cytoskeletal 4 OS = Mus musculus GN =Krt4 PE = 2 SV = 2 K2C73_MOUSE Keratin, type II cytoskeletal 73 OS = Musmusculus GN = Krt73 PE = 2 SV = 1 K1C17_MOUSE Keratin, type Icytoskeletal 17 OS = Mus musculus GN = Krt17 PE = 1 SV = 3 K22O_MOUSEKeratin, type II cytoskeletal 2 oral OS = Mus musculus GN = Krt76 PE = 2SV = 1 K2C79_MOUSE Keratin, type II cytoskeletal 79 OS = Mus musculus GN= Krt79 PE = 2 SV = 2 HBA_MOUSE Hemoglobin subunit alpha OS = Musmusculus GN = Hba PE = 1 SV = 2 AATC_MOUSE Aspartate aminotransferase,cytoplasmic OS = Mus musculus GN = Got1 PE = 1 SV = 2 K1C19_MOUSEKeratin, type I cytoskeletal 19 OS = Mus musculus GN = Krt19 PE = 2 SV =1 K1C42_MOUSE Keratin, type I cytoskeletal 42 OS = Mus musculus GN =Krt42 PE = 1 SV = 1 K2C1B_MOUSE Keratin, type II cytoskeletal 1b OS =Mus musculus GN = Krt77 PE = 1 SV = 1 K2C71_MOUSE Keratin, type IIcytoskeletal 71 OS = Mus musculus GN = Krt71 PE = 1 SV = 1 K2C74_MOUSEKeratin, type II cytoskeletal 74 OS = Mus musculus GN = Krt74 PE = 2 SV= 1 K2C8_MOUSE Keratin, type II cytoskeletal 8 OS = Mus musculus GN =Krt8 PE = 1 SV = 4 K1C27_MOUSE Keratin, type I cytoskeletal 27 OS = Musmusculus GN = Krt27 PE = 1 SV = 1 K2C72_MOUSE Keratin, type IIcytoskeletal 72 OS = Mus musculus GN = Krt72 PE = 2 SV = 1 FABP4_MOUSEFatty acid-binding protein, adipocyte OS = Mus musculus GN = Fabp4 PE =1 SV = 3 PTMS_MOUSE Parathymosin OS = Mus musculus GN = Ptms PE = 2 SV =3 1 K222P_MOUSE Keratin-like protein KRT222 OS = Mus musculus GN =Krt222 PE = 2 SV = 1 K22E_MOUSE Keratin, type II cytoskeletal 2epidermal OS = Mus musculus GN = Krt2 PE = 1 SV = 1 HBB2_MOUSEHemoglobin subunit beta-2 OS = Mus musculus GN = Hbb-b2 PE = 1 SV = 2TPIS_MOUSE Triosephosphate isomerase OS = Mus musculus GN = Tpi1 PE = 1SV = 3 FABPH_MOUSE Fatty acid-binding protein, heart OS = Mus musculusGN = Fabp3 PE = 1 SV = 5 BICD2_MOUSE Protein bicaudal D homolog 2 OS =Mus musculus GN = Bicd2 PE = 1 SV = 1 K2C1_PANTR Keratin, type IIcytoskeletal 1 OS = Pan troglodytes GN = KRT1 PE = 2 SV = 1 K2C1_HUMANKeratin, type II cytoskeletal 1 OS = Homo sapiens GN = KRT1 PE = 1 SV =5 K1C9_HUMAN Keratin, type I cytoskeletal 9 OS = Homo sapiens GN = KRT9PE = 1 SV = 2 K22E_HUMAN Keratin, type II cytoskeletal 2 epidermal OS =Homo sapiens GN = KRT2 PE = 1 SV = 1 K1C10_HUMAN Keratin, type Icytoskeletal 10 OS = Homo sapiens GN = KRT10 PE = 1 SV = 4 K1C10_CANFAKeratin, type I cytoskeletal 10 OS = Canis familiaris GN = KRT10 PE = 2SV = 1 K2C1_RAT Keratin, type II cytoskeletal 1 OS = Rattus norvegicusGN = Krt1 PE = 2 SV = 1 K22E_MOUSE Keratin, type II cytoskeletal 2epidermal OS = Mus musculus GN = Krt2 PE = 1 SV = 1 K1C14_HUMAN Keratin,type I cytoskeletal 14 OS = Homo sapiens GN = KRT14 PE = 1 SV = 3K1C10_BOVIN Keratin, type I cytoskeletal 10 OS = Bos taurus GN = KRT10PE = 3 SV = 1 K2C75_BOVIN Keratin, type II cytoskeletal 75 OS = Bostaurus GN = KRT75 PE = 2 SV = 1 TRYP_PIG Trypsin OS = Sus scrofa PE = 1SV = 1 K2C75_RAT Keratin, type II cytoskeletal 75 OS = Rattus norvegicusGN = Krt75 PE = 2 SV = 2 K2C6A_RAT Keratin, type II cytoskeletal 6A OS =Rattus norvegicus GN = Krt6a PE = 1 SV = 1 ALBU_MOUSE Serum albumin OS =Mus musculus GN = Alb PE = 1 SV = 3 K2C4_RAT Keratin, type IIcytoskeletal 4 OS = Rattus norvegicus GN = Krt4 PE = 2 SV = 1K2C73_HUMAN Keratin, type II cytoskeletal 73 OS = Homo sapiens GN =KRT73 PE = 1 SV = 1 K2C6A_HUMAN Keratin, type II cytoskeletal 6A OS =Homo sapiens GN = KRT6A PE = 1 SV = 3 K2C73_MOUSE Keratin, type IIcytoskeletal 73 OS = Mus musculus GN = Krt73 PE = 2 SV = 1 K2C1B_MOUSEKeratin, type II cytoskeletal 1b OS = Mus musculus GN = Krt77 PE = 1 SV= 1 K2C5_RAT Keratin, type II cytoskeletal 5 OS = Rattus norvegicus GN =Krt5 PE = 1 SV = 1 K2C6A_MOUSE Keratin, type II cytoskeletal 6A OS = Musmusculus GN = Krt6a PE = 2 SV = 3 K2C4_MOUSE Keratin, type IIcytoskeletal 4 OS = Mus musculus GN = Krt4 PE = 2 SV = 2 K22O_MOUSEKeratin, type II cytoskeletal 2 oral OS = Mus musculus GN = Krt76 PE = 2SV = 1 K2C3_HUMAN Keratin, type II cytoskeletal 3 OS = Homo sapiens GN =KRT3 PE = 1 SV = 2 K2C72_HUMAN Keratin, type II cytoskeletal 72 OS =Homo sapiens GN = KRT72 PE = 1 SV = 2 HBD_TARSY Hemoglobin subunit deltaOS = Tarsius syrichta GN = HBD PE = 2 SV = 2 3 K1C13_MOUSE Keratin, typeI cytoskeletal 13 OS = Mus musculus GN = Krt13 PE = 2 SV = 2 HBA_CRIGAHemoglobin subunit alpha OS = Cricetomys gambianus GN = HBA PE = 1 SV =2 TRY1_RAT Anionic trypsin-1 OS = Rattus norvegicus GN = Prss1 PE = 1 SV= 1 K2C8_MOUSE Keratin, type II cytoskeletal 8 OS = Mus musculus GN =Krt8 PE = 1 SV = 4 HBB_CALTO Hemoglobin subunit beta OS = Callicebustorquatus GN = HBB PE = 2 SV = 3 K1C3_XENLA Keratin, type I cytoskeletal47 kDa (Fragment) OS = Xenopus laevis GN = xk81b1 PE = 3 SV = 2K1C24_HUMAN Keratin, type I cytoskeletal 24 OS = Homo sapiens GN = KRT24PE = 1 SV = 1 K2C4_HUMAN Keratin, type II cytoskeletal 4 OS = Homosapiens GN = KRT4 PE = 1 SV = 4 AATC_HORSE Aspartate aminotransferase,cytoplasmic OS = Equus caballus GN = GOT1 PE = 1 SV = 2 K1C16_HUMANKeratin, type I cytoskeletal 16 OS = Homo sapiens GN = KRT16 PE = 1 SV =4 HBA_HUMAN Hemoglobin subunit alpha OS = Homo sapiens GN = HBA1 PE = 1SV = 2 HBA1_BUBBU Hemoglobin subunit alpha-1 OS = Bubalus bubalis PE = 2SV = 3 K1C42_RAT Keratin, type I cytoskeletal 42 OS = Rattus norvegicusGN = Krt42 PE = 2 SV = 1 K1C14_CHICK Keratin, type I cytoskeletal 14 OS= Gallus gallus GN = KRT14 PE = 2 SV = 1 HBA_MESAU Hemoglobin subunitalpha OS = Mesocricetus auratus GN = HBA PE = 1 SV = 1 UP03_PINHAUnknown protein 3 (Fragment) OS = Pinus halepensis PE = 1 SV = 1IGH1M_MOUSE Ig gamma-1 chain C region, membrane-bound form OS = Musmusculus GN = Ighg1 PE = 1 SV = 2 K2C5_PANTR Keratin, type IIcytoskeletal 5 OS = Pan troglodytes GN = KRT5 PE = 2 SV = 1 HBA_BISBOHemoglobin subunit alpha-I/II OS = Bison bonasus PE = 1 SV = 2 HBA_TALEUHemoglobin subunit alpha OS = Talpa europaea GN = HBA PE = 1 SV = 1HIR3_CHAGB Histone transcription regulator 3 homolog OS = Chaetomiumglobosum GN = HIR3 PE = 3 SV = 1 K1C10_HUMAN Keratin, type Icytoskeletal 10 OS = Homo sapiens GN = KRT10 PE = 1 SV = 4 K22E_HUMANKeratin, type II cytoskeletal 2 epidermal OS = Homo sapiens GN = KRT2 PE= 1 SV = 1 K1C9_HUMAN Keratin, type I cytoskeletal 9 OS = Homo sapiensGN = KRT9 PE = 1 SV = 2 K2C1_PANTR Keratin, type II cytoskeletal 1 OS =Pan troglodytes GN = KRT1 PE = 2 SV = 1 K2C1_HUMAN Keratin, type IIcytoskeletal 1 OS = Homo sapiens GN = KRT1 PE = 1 SV = 5 K1C10_CANFAKeratin, type I cytoskeletal 10 OS = Canis familiaris GN = KRT10 PE = 2SV = 1 K1C14_HUMAN Keratin, type I cytoskeletal 14 OS = Homo sapiens GN= KRT14 PE = 1 SV = 3 K2C5_PANTR Keratin, type II cytoskeletal 5 OS =Pan troglodytes GN = KRT5 PE = 2 SV = 1 K2C6A_HUMAN Keratin, type IIcytoskeletal 6A OS = Homo sapiens GN = KRT6A PE = 1 SV = 3 K1C16_HUMANKeratin, type I cytoskeletal 16 OS = Homo sapiens GN = KRT16 PE = 1 SV =4 TRYP_PIG Trypsin OS = Sus scrofa PE = 1 SV = 1 K2C79_HUMAN Keratin,type II cytoskeletal 79 OS = Homo sapiens GN = KRT79 PE = 1 SV = 1K1C15_HUMAN Keratin, type I cytoskeletal 15 OS = Homo sapiens GN = KRT15PE = 1 SV = 2 K1C15_SHEEP Keratin, type I cytoskeletal 15 OS = Ovisaries GN = KRT15 PE = 2 SV = 1 K1C10_BOVIN Keratin, type I cytoskeletal10 OS = Bos taurus GN = KRT10 PE = 3 SV = 1 K1C13_HUMAN Keratin, type Icytoskeletal 13 OS = Homo sapiens GN = KRT13 PE = 1 SV = 3 K2CO_CHICKKeratin, type II cytoskeletal cochleal OS = Gallus gallus PE = 2 SV = 1K2C1_RAT Keratin, type II cytoskeletal 1 OS = Rattus norvegicus GN =Krt1 PE = 2 SV = 1 K2C5_RAT Keratin, type II cytoskeletal 5 OS = Rattusnorvegicus GN = Krt5 PE = 1 SV = 1 K2C75_BOVIN Keratin, type IIcytoskeletal 75 OS = Bos taurus GN = KRT75 PE = 2 SV = 1 K2C7_BOVINKeratin, type II cytoskeletal 7 OS = Bos taurus GN = KRT7 PE = 2 SV = 1K2C5_BOVIN Keratin, type II cytoskeletal 5 OS = Bos taurus GN = KRT5 PE= 1 SV = 1 K2C6A_RAT Keratin, type II cytoskeletal 6A OS = Rattusnorvegicus GN = Krt6a PE = 1 SV = 1 K2C75_RAT Keratin, type IIcytoskeletal 75 OS = Rattus norvegicus GN = Krt75 PE = 2 SV = 2K2C73_MOUSE Keratin, type II cytoskeletal 73 OS = Mus musculus GN =Krt73 PE = 2 SV = 1 K22E_CANFA Keratin, type II cytoskeletal 2 epidermalOS = Canis familiaris GN = KRT2 PE = 2 SV = 1 K2C4_RAT Keratin, type IIcytoskeletal 4 OS = Rattus norvegicus GN = Krt4 PE = 2 SV = 1 5K1C3_XENLA Keratin, type I cytoskeletal 47 kDa (Fragment) OS = Xenopuslaevis GN = xk81b1 PE = 3 SV = 2 K22O_HUMAN Keratin, type IIcytoskeletal 2 oral OS = Homo sapiens GN = KRT76 PE = 1 SV = 1K2C3_RABIT Keratin, type II cytoskeletal 3 OS = Oryctolagus cuniculus GN= KRT3 PE = 2 SV = 1 K1C19_RAT Keratin, type I cytoskeletal 19 OS =Rattus norvegicus GN = Krt19 PE = 1 SV = 2 ALBU_MOUSE Serum albumin OS =Mus musculus GN = Alb PE = 1 SV = 3 K2C8_XENLA Keratin, type IIcytoskeletal 8 OS = Xenopus laevis PE = 2 SV = 1 K2C8_RAT Keratin, typeII cytoskeletal 8 OS = Rattus norvegicus GN = Krt8 PE = 1 SV = 3K22O_MOUSE Keratin, type II cytoskeletal 2 oral OS = Mus musculus GN =Krt76 PE = 2 SV = 1 K2C4_HUMAN Keratin, type II cytoskeletal 4 OS = Homosapiens GN = KRT4 PE = 1 SV = 4 TRY1_RAT Anionic trypsin-1 OS = Rattusnorvegicus GN = Prss1 PE = 1 SV = 1 K1C13_MOUSE Keratin, type Icytoskeletal 13 OS = Mus musculus GN = Krt13 PE = 2 SV = 2 K2C4_MOUSEKeratin, type II cytoskeletal 4 OS = Mus musculus GN = Krt4 PE = 2 SV =2 K2C8_DANRE Keratin, type II cytoskeletal 8 OS = Danio rerio GN = krt8PE = 1 SV = 1 KRT85_MOUSE Keratin, type II cuticular Hb5 OS = Musmusculus GN = Krt85 PE = 2 SV = 2 K2C80_BOVIN Keratin, type IIcytoskeletal 80 OS = Bos taurus GN = KRT80 PE = 2 SV = 1 K1C12_RABITKeratin, type I cytoskeletal 12 (Fragment) OS = Oryctolagus cuniculus GN= KRT12 PE = 2 SV = 1 K1C12_RAT Keratin, type I cytoskeletal 12 OS =Rattus norvegicus GN = Krt12 PE = 2 SV = 1 K2C1B_RAT Keratin, type IIcytoskeletal 1b OS = Rattus norvegicus GN = Krt77 PE = 2 SV = 1K1C1_XENLA Keratin, type I cytoskeletal 47 kDa OS = Xenopus laevis GN =xk81a1 PE = 2 SV = 1 K2C73_HUMAN Keratin, type II cytoskeletal 73 OS =Homo sapiens GN = KRT73 PE = 1 SV = 1 K1C24_HUMAN Keratin, type Icytoskeletal 24 OS = Homo sapiens GN = KRT24 PE = 1 SV = 1 K2C8_MOUSEKeratin, type II cytoskeletal 8 OS = Mus musculus GN = Krt8 PE = 1 SV =4 K2C78_HUMAN Keratin, type II cytoskeletal 78 OS = Homo sapiens GN =KRT78 PE = 1 SV = 2 K2C72_HUMAN Keratin, type II cytoskeletal 72 OS =Homo sapiens GN = KRT72 PE = 1 SV-2 K2C72_MOUSE Keratin, type IIcytoskeletal 72 OS = Mus musculus GN = Krt72 PE = 2 SV = 1 UP01_PINHAUnknown protein 1 (Fragment) OS = Pinus halepensis PE = 1 SV = 1K22E_MOUSE Keratin, type II cytoskeletal 2 epidermal OS = Mus musculusGN = Krt2 PE = 1 SV = 1 K1C14_CHICK Keratin, type I cytoskeletal 14 OS =Gallus gallus GN = KRT14 PE = 2 SV = 1 6 K1C27_HUMAN Keratin, type Icytoskeletal 27 OS = Homo sapiens GN = KRT27 PE = 1 SV = 1 UP03_PINHAUnknown protein 3 (Fragment) OS = Pinus halepensis PE = 1 SV = 1FDL30_ARATH Putative F-box/FBD/LRR-repeat protein At5g22610 OS =Arabidopsis thaliana GN = At5g22610 PE = 4 SV = 1 NDC80_CANAL Probablekinetochore protein NDC80 OS = Candida albicans GN = NDC80 PE = 3 SV = 1K1C10_HUMAN Keratin, type I cytoskeletal 10 OS = Homo sapiens GN = KRT10PE = 1 SV = 4 K22E_HUMAN Keratin, type II cytoskeletal 2 epidermal OS =Homo sapiens GN = KRT2 PE = 1 SV = 1 K2C1_PANTR Keratin, type IIcytoskeletal 1 OS = Pan troglodytes GN = KRT1 PE = 2 SV = 1 K1C10_CANFAKeratin, type I cytoskeletal 10 OS = Canis familiaris GN = KRT10 PE = 2SV = 1 K1C9_HUMAN Keratin, type I cytoskeletal 9 OS = Homo sapiens GN =KRT9 PE = 1 SV = 2 K2C1_HUMAN Keratin, type II cytoskeletal 1 OS = Homosapiens GN = KRT1 PE = 1 SV = 5 K2C75_BOVIN Keratin, type IIcytoskeletal 75 OS = Bos taurus GN = KRT75 PE = 2 SV = 1 K2C75_RATKeratin, type II cytoskeletal 75 OS = Rattus norvegicus GN = Krt75 PE =2 SV = 2 K2C1_RAT Keratin, type II cytoskeletal 1 OS = Rattus norvegicusGN = Krt1 PE = 2 SV = 1 K2C6A_RAT Keratin, type II cytoskeletal 6A OS =Rattus norvegicus GN = Krt6a PE = 1 SV = 1 K2C6A_HUMAN Keratin, type IIcytoskeletal 6A OS = Homo sapiens GN = KRT6A PE = 1 SV = 3 K2C5_BOVINKeratin, type II cytoskeletal 5 OS = Bos taurus GN = KRT5 PE = 1 SV = 1K2C5_MOUSE Keratin, type II cytoskeletal 5 OS = Mus musculus GN = Krt5PE = 2 SV = 1 K2C6A_MOUSE Keratin, type II cytoskeletal 6A OS = Musmusculus GN = Krt6a PE = 2 SV = 3 K2C79_BOVIN Keratin, type IIcytoskeletal 79 OS = Bos taurus GN = KRT79 PE = 2 SV = 1 ALBU_MOUSESerum albumin OS = Mus musculus GN = Alb PE = 1 SV = 3 K2C1_MOUSEKeratin, type II cytoskeletal 1 OS = Mus musculus GN = Krt1 PE = 1 SV =4 TRYP_PIG Trypsin OS = Sus scrofa PE = 1 SV = 1 K1C10_BOVIN Keratin,type I cytoskeletal 10 OS = Bos taurus GN = KRT10 PE = 3 SV = 1K22E_MOUSE Keratin, type II cytoskeletal 2 epidermal OS = Mus musculusGN = Krt2 PE = 1 SV = 1 HBA_SPAEH Hemoglobin subunit alpha OS = Spalaxleucodon ehrenbergi GN = HBA PE = 1 SV = 1 HBA_SPECI Hemoglobin subunitalpha OS = Spermophilus citellus GN = HBA PE = 1 SV = 1 K2C73_MOUSEKeratin, type II cytoskeletal 73 OS = Mus musculus GN = Krt73 PE = 2 SV= 1 K2C1B_MOUSE Keratin, type II cytoskeletal 1b OS = Mus musculus GN =Krt77 PE = 1 SV = 1 K2C4_RAT Keratin, type II cytoskeletal 4 OS = Rattusnorvegicus GN = Krt4 PE = 2 SV = 1 K2C73_HUMAN Keratin, type IIcytoskeletal 73 OS = Homo sapiens GN = KRT73 PE = 1 SV = 1 K2C5_PANTRKeratin, type II cytoskeletal 5 OS = Pan troglodytes GN = KRT5 PE = 2 SV= 1 8 TRY1_RAT Anionic trypsin-1 OS = Rattus norvegicus GN = Prss1 PE =1 SV = 1 K2C4_MOUSE Keratin, type II cytoskeletal 4 OS = Mus musculus GN= Krt4 PE = 2 SV = 2 K22O_MOUSE Keratin, type II cytoskeletal 2 oral OS= Mus musculus GN = Krt76 PE = 2 SV = 1 K2C3_HUMAN Keratin, type IIcytoskeletal 3 OS = Homo sapiens GN = KRT3 PE = 1 SV = 2 K2C72_HUMANKeratin, type II cytoskeletal 72 OS = Homo sapiens GN = KRT72 PE = 1 SV= 2 K2C72_MOUSE Keratin, type II cytoskeletal 72 OS = Mus musculus GN =Krt72 PE = 2 SV = 1 HBA_MOUSE Hemoglobin subunit alpha OS = Mus musculusGN = Hba PE = 1 SV = 2 K1C14_HUMAN Keratin, type I cytoskeletal 14 OS =Homo sapiens GN = KRT14 PE = 1 SV = 3 K1C3_XENLA Keratin, type Icytoskeletal 47 kDa (Fragment) OS = Xenopus laevis GN = xk81b1 PE = 3 SV= 2 K1C1_XENLA Keratin, type I cytoskeletal 47 kDa OS = Xenopus laevisGN = xk81a1 PE = 2 SV = 1 K1C12_RABIT Keratin, type I cytoskeletal 12(Fragment) OS = Oryctolagus cuniculus GN = KRT12 PE = 2 SV = 1 K1C12_RATKeratin, type I cytoskeletal 12 OS = Rattus norvegicus GN = Krt12 PE = 2SV = 1 K2C8_MOUSE Keratin, type II cytoskeletal 8 OS = Mus musculus GN =Krt8 PE = 1 SV = 4 ALBU_FELCA Serum albumin OS = Felis silvestris catusGN = ALB PE = 1 SV = 1 K2C4_HUMAN Keratin, type II cytoskeletal 4 OS =Homo sapiens GN = KRT4 PE = 1 SV = 4 HBB_CALTO Hemoglobin subunit betaOS = Callicebus torquatus GN = HBB PE = 2 SV = 3 K1C25_BOVIN Keratin,type I cytoskeletal 25 OS = Bos taurus GN = KRT25 PE = 2 SV = 1HBA_MESAU Hemoglobin subunit alpha OS = Mesocricetus auratus GN = HBA PE= 1 SV = 1 K2C8_BOVIN Keratin, type II cytoskeletal 8 OS = Bos taurus GN= KRT8 PE = 2 SV = 3 K2C8_RAT Keratin, type II cytoskeletal 8 OS =Rattus norvegicus GN = Krt8 PE = 1 SV = 3 MATK_ADELA Maturase K OS =Adesmia lanata GN = matK PE = 3 SV = 1 HBA_BISBO Hemoglobin subunitalpha-I/II OS = Bison bonasus PE = 1 SV = 2 HBA_TALEU Hemoglobin subunitalpha OS = Talpa europaea GN = HBA PE = 1 SV = 1 K1C10_HUMAN Keratin,type I cytoskeletal 10 OS = Homo sapiens GN = KRT10 PE = 1 SV = 4K1C9_HUMAN Keratin, type I cytoskeletal 9 OS = Homo sapiens GN = KRT9 PE= 1 SV = 2 K1C10_CANFA Keratin, type I cytoskeletal 10 OS = Canisfamiliaris GN = KRT10 PE = 2 SV = 1 K22E_HUMAN Keratin, type IIcytoskeletal 2 epidermal OS = Homo sapiens GN = KRT2 PE = 1 SV = 1K2C1_HUMAN Keratin, type II cytoskeletal 1 OS = Homo sapiens GN = KRT1PE = 1 SV = 5 K2C1_PANTR Keratin, type II cytoskeletal 1 OS = Pantroglodytes GN = KRT1 PE = 2 SV = 1 CASQ1_MOUSE Calsequestrin-1 OS = Musmusculus GN = Casq1 PE = 2 SV = 2 K2C6A_HUMAN Keratin, type IIcytoskeletal 6A OS = Homo sapiens GN = KRT6A PE = 1 SV = 3 HBB_GORGOHemoglobin subunit beta OS = Gorilla gorilla gorilla GN = HBB PE = 1 SV= 2 K2C5_HUMAN Keratin, type II cytoskeletal 5 OS = Homo sapiens GN =KRT5 PE = 1 SV = 3 K2C1_RAT Keratin, type II cytoskeletal 1 OS = Rattusnorvegicus GN = Krt1 PE = 2 SV = 1 K1C10_BOVIN Keratin, type Icytoskeletal 10 OS = Bos taurus GN = KRT10 PE = 3 SV = 1 HBB_MELMEHemoglobin subunit beta OS = Meles meles GN = HBB PE = 1 SV = 1HBB_CALTO Hemoglobin subunit beta OS = Callicebus torquatus GN = HBB PE= 2 SV = 3 HBA_HUMAN Hemoglobin subunit alpha OS = Homo sapiens GN =HBA1 PE = 1 SV = 2 ALBU_RAT Serum albumin OS = Rattus norvegicus GN =Alb PE = 1 SV = 2 ALBU_FELCA Serum albumin OS = Felis silvestris catusGN = ALB PE = 1 SV = 1 K2C6A_RAT Keratin, type II cytoskeletal 6A OS =Rattus norvegicus GN = Krt6a PE = 1 SV = 1 ALBU_HUMAN Serum albumin OS =Homo sapiens GN = ALB PE = 1 SV = 2 TRYP_PIG Trypsin OS = Sus scrofa PE= 1 SV = 1 K2C75_BOVIN Keratin, type II cytoskeletal 75 OS = Bos taurusGN = KRT75 PE = 2 SV = 1 MLE1_HUMAN Myosin light chain 1, skeletalmuscle isoform OS = Homo sapiens GN = MYL1 PE = 1 SV = 3 ACTS_ATRMMActin, alpha skeletal muscle OS = Atractaspis microlepidotamicrolepidota GN = ACTA1 PE = 2 SV = 1 ACT2_MOLOC Actin, muscle-type OS=Molgula oculata PE = 3 SV = 1 HBB_NASNA Hemoglobin subunit beta OS =Nasua nasua GN = HBB PE = 1 SV = 1 HBB_CANFA Hemoglobin subunit beta OS= Canis familiaris GN = HBB PE = 1 SV = 1 10 HBA_MESAU Hemoglobinsubunit alpha OS = Mesocricetus auratus GN = HBA PE = 1 SV = 1K2C5_BOVIN Keratin, type II cytoskeletal 5 OS = Bos taurus GN = KRT5 PE= 1 SV = 1 K2C6A_MOUSE Keratin, type II cytoskeletal 6A OS = Musmusculus GN = Krt6a PE = 2 SV = 3 HBB_MACMU Hemoglobin subunit beta OS =Macaca mulatta GN = HBB PE = 1 SV = 1 HBB_SAISC Hemoglobin subunit betaOS = Saimiri sciureus GN = HBB PE = 1 SV = 2 TRY1_RAT Anionic trypsin-1OS = Rattus norvegicus GN = Prss1 PE = 1 SV = 1 K2C8_MOUSE Keratin, typeII cytoskeletal 8 OS = Mus musculus GN = Krt8 PE = 1 SV = 4 K2C1B_RATKeratin, type II cytoskeletal 1b OS = Rattus norvegicus GN = Krt77 PE =2 SV = 1 MDHM_BOVIN Malate dehydrogenase, mitochondrial OS = Bos taurusGN = MDH2 PE = 2 SV = 1 HBA_CRIGA Hemoglobin subunit alpha OS =Cricetomys gambianus GN = HBA PE = 1 SV = 2 HBA_SPECI Hemoglobin subunitalpha OS = Spermophilus citellus GN = HBA PE = 1 SV = 1 HBB_PAGLAHemoglobin subunit beta OS = Paguma larvata GN = HBB PE = 1 SV = 1HBB_MYOVE Hemoglobin subunit beta OS = Myotis velifer GN = HBB PE = 1 SV= 1 K1C1_XENLA Keratin, type I cytoskeletal 47 kDa OS = Xenopus laevisGN = xk81a1 PE = 2 SV = 1 ACT18_DICDI Actin-18 OS = Dictyosteliumdiscoideum GN = act18 PE = 3 SV = 3 K1C24_HUMAN Keratin, type Icytoskeletal 24 OS = Homo sapiens GN = KRT24 PE = 1 SV = 1 ACT1_ARTSXActin, clone 205 OS = Artemia sp. PE = 2 SV = 1 ACTC_BRAFL Actin,cytoplasmic OS = Branchiostoma floridae PE = 2 SV = 1 ACTB3_FUGRU Actin,cytoplasmic 3 OS = Fugu rubripes GN = actbc PE = 2 SV = 1 ACT_THELAActin OS = Thermomyces lanuginosus PE = 3 SV = 1 ACT1_SACKO Actin-1 OS =Saccoglossus kowalevskii PE = 2 SV = 1 ACT1_LYTPI Actin, cytoskeletal 1OS = Lytechinus pictus PE = 2 SV = 1 ACTC_BRALA Actin, cytoplasmic OS =Branchiostoma lanceolatum PE = 2 SV = 1 ACT_CANGA Actin OS = Candidaglabrata GN = ACT1 PE = 3 SV = 1 ACT_KLULA Actin OS = Kluyveromyceslactis GN = ACT PE = 3 SV = 2 ACT_PHARH Actin OS = Phaffia rhodozyma PE= 3 SV = 1 ACT_CANAL Actin OS = Candida albicans GN = ACT1 PE = 3 SV = 1ACT_SCHPO Actin OS = Schizosaccharomyces pombe GN = act1 PE = 1 SV = 111 ACTBL_HUMAN Beta-actin-like protein 2 OS = Homo sapiens GN = ACTBL2PE = 1 SV = 2 A26CA_HUMAN ANKRD26-like family C member 1A OS = Homosapiens GN = A26C1A PE = 1 SV = 3 K2C4_RAT Keratin, type II cytoskeletal4 OS = Rattus norvegicus GN = Krt4 PE = 2 SV = 1 K2C73_HUMAN Keratin,type II cytoskeletal 73 OS = Homo sapiens GN = KRT73 PE = 1 SV = 1HBA_MACFA Hemoglobin subunit alpha-A/Q/R/T OS = Macaca fascicularis PE =1 SV = 1 HBD_GORGO Hemoglobin subunit delta OS = Gorilla gorilla gorillaGN = HBD PE = 1 SV = 2 MDHM_YEAST Malate dehydrogenase, mitochondrial OS= Saccharomyces cerevisiae GN = MDH1 PE = 1 SV = 2 HBA_LAMGL Hemoglobinsubunit alpha OS = Lama glama GN = HBA PE = 1 SV = 1 HBB_RABITHemoglobin subunit beta-1/2 OS = Oryctolagus cuniculus GN = HBB1 PE = 1SV = 2 HBD_ELEMA Hemoglobin subunit delta OS = Elephas maximus GN = HBDPE = 2 SV = 3 FIBB_HUMAN Fibrinogen beta chain OS = Homo sapiens GN =FGB PE = 1 SV = 2 HBD_COLPO Hemoglobin subunit delta OS = Colobuspolykomos GN = HBD PE = 2 SV = 2 K2C4_MOUSE Keratin, type IIcytoskeletal 4 OS = Mus musculus GN = Krt4 PE = 2 SV = 2 K22O_MOUSEKeratin, type II cytoskeletal 2 oral OS = Mus musculus GN = Krt76 PE = 2SV = 1 K2C3_RABIT Keratin, type II cytoskeletal 3 OS = Oryctolaguscuniculus GN = KRT3 PE = 2 SV = 1 TPISA_DANRE Triosephosphate isomeraseA OS = Danio rerio GN = tpi1a PE = 2 SV = 1 TPIS_ASPOR Triosephosphateisomerase OS = Aspergillus oryzae GN = tpiA PE = 2 SV = 1 TPIS_HERARTriosephosphate isomerase OS = Herminiimonas arsenicoxydans GN = tpiA PE= 3 SV = 1 VIM4_XENLA Vimentin-4 OS = Xenopus laevis GN = vim4 PE = 2 SV= 1 VIME_BOVIN Vimentin OS = Bos taurus GN = VIM PE = 1 SV = 3MATK_ADELA Maturase K OS = Adesmia lanata GN = matK PE = 3 SV = 1CASA1_BOVIN Alpha-S1-casein OS = Bos taurus GN = CSN1S1 PE = 1 SV = 2MLE1_CHICK Myosin light chain 1, skeletal muscle isoform OS = Gallusgallus PE = 1 SV = 3 HBA_ARAAR Hemoglobin subunit alpha-A OS = Araararauna GN = HBAA PE = 1 SV = 2 PYRB_AERHH Aspartatecarbamoyltransferase OS = Aeromonas hydrophila subsp. hydrophila (strainATCC 7966/NCIB 9240) GN = pyrB PE = 3 SV = 2 ATPB_BOVIN ATP synthasesubunit beta, mitochondrial OS = Bos taurus GN = ATP5B PE = 1 SV = 2Q8BGZ7_MOUSE 10 days neonate skin cDNA, RIKEN full-length enrichedlibrary, clone: 4732475I03 product: CYTOKERATIN homolog (10 days neonateskin cDNA, RIKEN full-length enriched library, clone: 4732468K03product: CYTOKERATIN TYPE II homolog) (6 days neonate skin cD K2C1_MOUSEKeratin, type II cytoskeletal 1 (Cytokeratin-1) (CK-1) (Keratin-1) (K1)(67 kDa cytokeratin). - Mus musculus (Mouse). Q9D2K8_MOUSE 0 day neonatehead cDNA, RIKEN full-length enriched library, clone: 4833436C19product: keratin complex 2, basic, gene 1, full insert sequence. - Musmusculus (Mouse). Q32P04_MOUSE Krt2-5 protein (Fragment). - Mus musculus(Mouse). K2C6A_MOUSE Keratin, type II cytoskeletal 6A (Cytokeratin-6A)(CK 6A) (K6a keratin) (Keratin-6 alpha) (mK6-alpha). - Mus musculus(Mouse). Q80VP7_MOUSE Hypothetical protein MGC54654. - Mus musculus(Mouse). Q8BIS2_MOUSE 10 days neonate skin cDNA, RIKEN full-lengthenriched library, clone: 4732456N10 product: similar to KERATIN, TYPE IICYTOSKELETAL 6D (CYTOKERATIN 6D) (CK 6D) (K6D KERATIN). - Mus musculus(Mouse). Q9CXH5_MOUSE 17 days embryo head cDNA, RIKEN full-lengthenriched library, clone: 3300001P16 product: hemoglobin, beta adultmajor chain, full insert sequence. - Mus musculus (Mouse). 13 KRMSE1keratin, 59K type I cytoskeletal - mouse Q6NXH9_MOUSE Type II keratinKb36. - Mus musculus (Mouse). K2C1B_MOUSE Keratin, type II cytoskeletal1b (Type II keratin Kb39) (Embryonic type II keratin-1). - Mus musculus(Mouse). Q6IFZ9_MOUSE Type II keratin Kb37. - Mus musculus (Mouse).Q3TTY5_MOUSE 10 days neonate skin cDNA, RIKEN full-length enrichedlibrary, clone: 4732404G19 product: keratin complex 2, basic, gene 17,full insert sequence (10 days neonate skin cDNA, RIKEN full-lengthenriched library, clone: 4732426A12 product: keratin complexQ6IFT3_MOUSE Keratin Kb40. - Mus musculus (Mouse). Q3UV17_MOUSE Adultfemale vagina cDNA, RIKEN full-length enriched library, clone:9930024P18 product: similar to Keratin 2p. - Mus musculus (Mouse).I59009 epidermal keratin subunit II - mouse HAMS hemoglobin alphachains - mouse K2C4_MOUSE Keratin, type II cytoskeletal 4(Cytokeratin-4) (CK-4) (Keratin-4) (K4) (Cytoskeletal 57 kDa keratin). -Mus musculus (Mouse). 14 Q6IME9_MOUSE Type-II keratin Kb35. - Musmusculus (Mouse). K2C8_MOUSE Keratin, type II cytoskeletal 8(Cytokeratin-8) (CK-8) (Keratin-8) (K8) (Cytokeratin endo A). - Musmusculus (Mouse). Q3TV03_MOUSE Adult male stomach cDNA, RIKENfull-length enriched library, clone: 2210414C06 product: albumin 1, fullinsert sequence. - Mus musculus (Mouse). Q9Z1R9_MOUSE Trypsinogen 16(Protease, serine, 1). - Mus musculus (Mouse). JQ0028 cytokeratin 19 -mouse Q2M1G8_MOUSE 2410039E07Rik protein (Fragment). - Mus musculus(Mouse). AAH11074 BC011074 NID: - Mus musculus KRMSE1 keratin, 59K typeI cytoskeletal - mouse Q9CY54_MOUSE 13 days embryo liver cDNA, RIKENfull-length enriched library, clone: 2500004H04 product: hemoglobin,beta adult major chain, full insert sequence. - Mus musculus (Mouse).HAMS hemoglobin alpha chains - mouse Q9CY06_MOUSE 13 days embryo livercDNA, RIKEN full-length enriched library, clone: 2510040P05 product:hemoglobin, beta adult major chain, full insert sequence. - Mus musculus(Mouse). AAH11074 BC011074 NID: - Mus musculus HBMS hemoglobin betamajor chain - mouse Q3TTY5_MOUSE 10 days neonate skin cDNA, RIKENfull-length enriched library, clone: 4732404G19 product: keratin complex2, basic, gene 17, full insert sequence (10 days neonate skin cDNA,RIKEN full-length enriched library, clone: 4732426A12 product: keratincomplex Q3UJH8_MOUSE 16 days embryo heart cDNA, RIKEN full-lengthenriched library, clone: I920001F12 product: glutamate oxaloacetatetransaminase 1, soluble, full insert sequence. - Mus musculus (Mouse).Q8BGZ7_MOUSE 10 days neonate skin cDNA, RIKEN full-length enrichedlibrary, clone: 4732475I03 product: CYTOKERATIN homolog (10 days neonateskin cDNA, RIKEN full-length enriched library, clone: 4732468K03product: CYTOKERATIN TYPE II homolog) (6 days neonate skin cD 16K1C17_MOUSE Keratin, type I cytoskeletal 17 (Cytokeratin-17) (CK-17)(Keratin-17) (K17). - Mus musculus (Mouse). Q9R0S6_MOUSE Beta-1-globin(Fragment). - Mus musculus (Mouse). K2C1_MOUSE Keratin, type IIcytoskeletal 1 (Cytokeratin-1) (CK-1) (Keratin-1) (Kl) (67 kDacytokeratin). - Mus musculus (Mouse). Q9D2K8_MOUSE 0 day neonate headcDNA, RIKEN full-length enriched library, clone: 4833436C19 product:keratin complex 2, basic, gene 1, full insert sequence. - Mus musculus(Mouse). A55682 keratin 13, type I cytoskeletal - mouse Q32P04_MOUSEKrt2-5 protein (Fragment). - Mus musculus (Mouse). CAA33084 MMHFABPNID: - Mus musculus Q80VP7_MOUSE Hypothetical protein MGC54654. - Musmusculus (Mouse). Q8BIS2_MOUSE 10 days neonate skin cDNA, RIKENfull-length enriched library, clone: 4732456N10 product: similar toKERATIN, TYPE II CYTOSKELETAL 6D (CYTOKERATIN 6D) (CK 6D) (K6DKERATIN). - Mus musculus (Mouse). Q6NXH9_M Type II keratin Kb36. - Musmusculus (Mouse). 17 OUSE K1C15_MOUSE Keratin, type I cytoskeletal 15(Cytokeratin-15) (CK-15) (Keratin-15) (K15). - Mus musculus (Mouse).FABPH_MOUSE Fatty acid-binding protein, heart (H-FABP) (Heart-type fattyacid- binding protein) (Mammary-derived growth inhibitor) (MDGI). - Musmusculus (Mouse). Q3UV17_MOUSE Adult female vagina cDNA, RIKENfull-length enriched library, clone: 9930024P18 product: similar toKeratin 2p. - Mus musculus (Mouse). K2C4_MOUSE Keratin, type IIcytoskeletal 4 (Cytokeratin-4) (CK-4) (Keratin-4) (K4) (Cytoskeletal 57kDa keratin). - Mus musculus (Mouse). Q6IME9_MOUSE Type-II keratinKb35. - Mus musculus (Mouse). Q6IFT3_MOUSE Keratin Kb40. - Mus musculus(Mouse). Q8VCW2_MOUSE RIKEN cDNA 4631426H08. - Mus musculus (Mouse).Q2M1G8_MOUSE 2410039E07Rik protein (Fragment). - Mus musculus (Mouse).FABPA_MO Fatty acid-binding protein, adipocyte (AFABP) (Adipocytelipid-binding protein) (ALBP) (A-FABP) (P2 adipocyte protein) (Myelin P2protein homolog) (3T3-L1 lipid-binding protein) (422 protein) (P15). -18 USE Mus musculus (Mouse). I59009 epidermal keratin subunit II - mouseQ3TV03_MOUSE Adult male stomach cDNA, RIKEN full-length enrichedlibrary, clone: 2210414C06 product: albumin 1, full insert sequence. -Mus musculus (Mouse). DEMSMC malate dehydrogenase (EC 1.1.1.37),cytosolic - mouse JQ0028 cytokeratin 19 - mouse Q3UDM1_MOUSE Bone marrowmacrophage cDNA, RIKEN full-length enriched library, clone: G530119K21product: ATP-binding cassette, sub-family C (CFTR/MRP), member 1, fullinsert sequence. (Fragment). - Mus musculus (Mouse). Q3UBW7_MOUSE Bonemarrow macrophage cDNA, RIKEN full-length enriched library, clone:I830015F18 product: transferrin, full insert sequence. - Mus musculus(Mouse). K1C10_MOUSE Keratin, type I cytoskeletal 10 OS = Mus musculusGN = Krt10 PE = 1 SV = 3 K2C75_MOUSE Keratin, type II cytoskeletal 75 OS= Mus musculus GN = Krt75 PE = 1 SV = 1 ALBU_MOUSE Serum albumin OS =Mus musculus GN = Alb PE = 1 SV = 3 K2C5_MOUSE Keratin, type IIcytoskeletal 5 OS = Mus musculus GN = Krt5 PE = 2 SV = 1 K2C1_MOUSEKeratin, type II cytoskeletal 1 OS = Mus musculus GN = Krt1 PE = 1 SV =4 K2C6A_MOUSE Keratin, type II cytoskeletal 6A OS = Mus musculus GN =Krt6a PE = 2 SV = 3 K2C6B_MOUSE Keratin, type II cytoskeletal 6B OS =Mus musculus GN = Krt6b PE = 2 SV = 3 K2C79_MOUSE Keratin, type IIcytoskeletal 79 OS = Mus musculus GN = Krt79 PE = 2 SV = 2 K1C13_MOUSEKeratin, type I cytoskeletal 13 OS = Mus musculus GN = Krt13 PE = 2 SV =2 AATC_MOUSE Aspartate aminotransferase, cytoplasmic OS = Mus musculusGN = Got1 PE = 1 SV = 2 K1C14_MOUSE Keratin, type I cytoskeletal 14 OS =Mus musculus GN = Krt14 PE = 1 SV = 2 K1C17_MOUSE Keratin, type Icytoskeletal 17 OS = Mus musculus GN = Krt17 PE = 1 SV = 3 K2C73_MOUSEKeratin, type II cytoskeletal 73 OS = Mus musculus GN = Krt73 PE = 2 SV= 1 HBB1_MOUSE Hemoglobin subunit beta-1 OS = Mus musculus GN = Hbb-b1PE = 1 SV = 2 K2C71_MOUSE Keratin; type II cytoskeletal 71 OS = Musmusculus GN = Krt71 PE = 1 SV = 1 K2C4_MOUSE Keratin, type IIcytoskeletal 4 OS = Mus musculus GN = Krt4 PE = 2 SV = 2 K22O_MOUSEKeratin, type II cytoskeletal 2 oral OS = Mus musculus GN = Krt76 PE = 2SV = 1 K2C72_MOUSE Keratin, type II cytoskeletal 72 OS = Mus musculus GN= Krt72 PE = 2 SV = 1 K2C8_MOUSE Keratin, type II cytoskeletal 8 OS =Mus musculus GN = Krt8 PE = 1 SV = 4 FABPH_MOUSE Fatty acid-bindingprotein, heart OS = Mus musculus GN = Fabp3 PE = 1 SV = 5 SOX1_MOUSESOX-1 protein OS = Mus musculus GN = Sox1 PE = 2 SV = 1 SPA3M_MOUSESerine protease inhibitor A3M OS = Mus musculus GN = Serpina3m PE = 1 SV= 1 TPIS_MOUSE Triosephosphate isomerase OS = Mus musculus GN = Tpi1 PE= 1 SV = 3 K1C15_MOUSE Keratin, type I cytoskeletal 15 OS = Mus musculusGN = Krt15 PE = 1 SV = 2 K1C16_MOUSE Keratin, type I cytoskeletal 16 OS= Mus musculus GN = Krt16 PE = 1 SV = 3 K22E_MOUSE Keratin, type IIcytoskeletal 2 epidermal OS = Mus musculus GN = Krt2 PE = 1 SV = 1 20DNJC1_MOUSE DnaJ homolog subfamily C member 1 OS = Mus musculus GN =Dnajc1 PE = 1 SV = 1 HBA_MOUSE Hemoglobin subunit alpha OS = Musmusculus GN = Hba PE = 1 SV = 2 ARVC_MOUSE Armadillo repeat proteindeleted in velo-cardio-facial syndrome homolog OS = Mus musculus GN =Arvcf PE = 1 SV = 2 VIME_MOUSE Vimentin OS = Mus musculus GN = Vim PE =1 SV = 3 KRT83_MOUSE Keratin, type II cuticular Hb3 OS = Mus musculus GN= Krt83 PE = 2 SV = 2 KRT82_MOUSE Keratin, type II cuticular Hb2 OS =Mus musculus GN = Krt82 PE = 2 SV = 1 A1AT1_MOUSE Alpha-1-antitrypsin1-1 OS = Mus musculus GN = Serpina1a PE = 1 SV = 4 SODC_MOUSE Superoxidedismutase [Cu—Zn] OS = Mus musculus GN = Sod1 PE = 1 SV = 2 AS250_MOUSE250 kDa substrate of Akt OS = Mus musculus GN = Kiaa1272 PE = 1 SV = 2K1C25_MOUSE Keratin, type I cytoskeletal 25 OS = Mus musculus GN = Krt25PE = 1 SV = 1 HBB1_MOUSE Hemoglobin subunit beta-1 OS = Mus musculus GN= Hbb-b1 PE = 1 SV = 2 K1C10_MOUSE Keratin, type I cytoskeletal 10 OS =Mus musculus GN = Krt10 PE = 1 SV = 3 K2C75_MOUSE Keratin, type IIcytoskeletal 75 OS = Mus musculus GN = Krt75 PE = 1 SV = 1 K2C1_MOUSEKeratin, type II cytoskeletal 1 OS = Mus musculus GN = Krt1 PE = 1 SV =4 K2C5_MOUSE Keratin, type II cytoskeletal 5 OS = Mus musculus GN = Krt5PE = 2 SV = 1 ALBU_MOUSE Serum albumin OS = Mus musculus GN = Alb PE = 1SV = 3 K2C79_MOUSE Keratin, type II cytoskeletal 79 OS = Mus musculus GN= Krt79 PE = 2 SV = 2 K1C42_MOUSE Keratin, type I cytoskeletal 42 OS =Mus musculus GN = Krt42 PE = 1 SV = 1 HBA_MOUSE Hemoglobin subunit alphaOS = Mus musculus GN = Hba PE = 1 SV = 2 K2C73_MOUSE Keratin, type IIcytoskeletal 73 OS = Mus musculus GN = Krt73 PE = 2 SV = 1 K1C15_MOUSEKeratin, type I cytoskeletal 15 OS = Mus musculus GN = Krt15 PE = 1 SV =2 K2C1B_MOUSE Keratin, type II cytoskeletal 1b OS = Mus musculus GN =Krt77 PE = 1 SV = 1 K1C27_MOUSE Keratin, type I cytoskeletal 27 OS = Musmusculus GN = Krt27 PE = 1 SV = 1 K2C4_MOUSE Keratin; type IIcytoskeletal 4 OS = Mus musculus GN = Krt4 PE = 2 SV = 2 K22O_MOUSEKeratin, type II cytoskeletal 2 oral OS = Mus musculus GN = Krt76 PE = 2SV = 1 K2C72_MOUSE Keratin, type II cytoskeletal 72 OS = Mus musculus GN= Krt72 PE = 2 SV = 1 K1C19_MOUSE Keratin, type I cytoskeletal 19 OS =Mus musculus GN = Krt19 PE = 2 SV = 1 K1C17_MOUSE Keratin, type Icytoskeletal 17 OS = Mus musculus GN = Krt17 PE = 1 SV = 3 K22E_MOUSEKeratin, type II cytoskeletal 2 epidermal OS = Mus musculus GN = Krt2 PE= 1 SV = 1 SYTC_MOUSE Threonyl-tRNA synthetase, cytoplasmic OS = Musmusculus GN = Tars PE = 1 SV = 2 HBE_MOUSE Hemoglobin subunit epsilon-Y2OS = Mus musculus GN = Hbb-y PE = 1 SV = 2 FTSJ2_MOUSE FtsJmethyltransferase domain-containing protein 2 OS = Mus musculus GN =Ftsjd2 PE = 1 SV = 1 DPOLA_MOUSE DNA polymerase alpha catalytic subunitOS = Mus musculus GN = Pola1 PE = 1 SV = 2 HBB1_MOUSE Mass: 15830 Score:216 Queries matched: 10 emPAI: 2.20 Hemoglobin subunit beta-1 OS = Musmusculus GN = Hbb-b1 PE = 1 SV = 2 Check to include this hit in errortolerant search or archive report HAMS hemoglobin alpha chains - mouseQ9CY12_MOUSE 13 days embryo liver cDNA, RIKEN full-length enrichedlibrary, clone: 2510039D09 product: hemoglobin, beta adult major chain,full insert sequence. - Mus musculus (Mouse). Q8BGZ7_MOUSE 10 daysneonate skin cDNA, RIKEN full-length enriched library, clone: 4732475I03product: CYTOKERATIN homolog (10 days neonate skin cDNA, RIKENfull-length enriched library, clone: 4732468K03 product: CYTOKERATINTYPE II homolog) (6 days neonate skin cD Q32P04_MOUSE Krt2-5 protein(Fragment). - Mus musculus (Mouse). Q3TV03_MOUSE Adult male stomachcDNA, RIKEN full-length enriched library, clone: 2210414C06 product:albumin 1, full insert sequence. - Mus musculus (Mouse). KRMSE1 keratin,59K type I cytoskeletal - mouse K1C15_MOUSE Keratin, type I cytoskeletal15 (Cytokeratin-15) (CK-15) (Keratin-15) (K15). - Mus musculus (Mouse).Q8BIS2_MOUSE 10 days neonate skin cDNA, RIKEN full-length enrichedlibrary, clone: 4732456N10 product: similar to KERATIN, TYPE IICYTOSKELETAL 6D (CYTOKERATIN 6D) (CK 6D) (K6D KERATIN). - Mus musculus(Mouse). Q8BGZ7_MOUSE 10 days neonate skin cDNA, RIKEN full-lengthenriched library, clone: 4732475I03 product: CYTOKERATIN homolog (10days neonate skin cDNA, RIKEN full-length enriched library, clone:4732468K03 product: CYTOKERATIN TYPE II homolog) (6 days neonate skin cDK2C6A_MOUSE Keratin, type II cytoskeletal 6A (Cytokeratin-6A). (CK 6A)(K6a keratin) (Keratin-6 alpha) (mK6-alpha). - Mus musculus (Mouse).Q32P04_MOUSE Krt2-5 protein (Fragment). - Mus musculus (Mouse).Q8K2E4_MOUSE CDNA sequence BC031593. - Mus musculus (Mouse). K2C1_MOUSEKeratin, type II cytoskeletal 1 (Cytokeratin-1) (CK-1) (Keratin-1) (K1)(67 kDa cytokeratin). - Mus musculus (Mouse). Q9D2K8_MOUSE 0 day neonatehead cDNA, RIKEN full-length enriched library, clone: 4833436C19product: keratin complex 2, basic, gene 1, full insert sequence. - Musmusculus (Mouse). Q8BIS2_MOUSE 10 days neonate skin cDNA, RIKENfull-length enriched library, clone: 4732456N10 product: similar toKERATIN, TYPE II CYTOSKELETAL 6D (CYTOKERATIN 6D) (CK 6D) (K6DKERATIN). - Mus musculus (Mouse). Q80VP7_MOUSE Hypothetical proteinMGC54654. - Mus musculus (Mouse). Q6NXH9_MOUSE Type II keratin Kb36. -Mus musculus (Mouse). Q6IFT3_MO Keratin Kb40. - Mus musculus (Mouse). 25USE K2C1B_MOUSE Keratin, type II cytoskeletal 1b (Type II keratin Kb39)(Embryonic type II keratin-1). - Mus musculus (Mouse). Q3UV17_MOUSEAdult female vagina cDNA, RIKEN full-length enriched library, clone:9930024P18 product: similar to Keratin 2p. - Mus musculus (Mouse).K2C4_MOUSE Keratin, type II cytoskeletal 4 (Cytokeratin-4) (CK-4)(Keratin-4) (K4) (Cytoskeletal 57 kDa keratin). - Mus musculus (Mouse).AAH11074 BC011074 NID: - Mus musculus BAA85657 AB033744 NID: - Musmusculus Q6IFZ9_MOUSE Type II keratin Kb37. - Mus musculus (Mouse).I59009 epidermal keratin subunit II - mouse K1C17_MOUSE Keratin, type Icytoskeletal 17 (Cytokeratin-17) (CK-17) (Keratin-17) (K17). - Musmusculus (Mouse). Q6IFX2_MOUSE Type I keratin KA22. - Mus musculus(Mouse). Q6IME9_MOUSE Type-II keratin Kb35. - Mus musculus (Mouse).BAE40567 AK168726 NID: - Mus musculus AAD01692 AF021836 NID: - Musmusculus Q3USS4 Adult male corpora quadrigemina cDNA, RIKEN full-lengthenriched library, clone: B230315D14 product: glial fibrillary acidicprotein, full insert sequence. - Mus musculus (Mouse). 26 _MOUSEQ3TTY5_MOUSE 10 days neonate skin cDNA, RIKEN full-length enrichedlibrary, clone: 4732404G19 product: keratin complex 2, basic, gene 17,full insert sequence (10 days neonate skin cDNA, RIKEN full-lengthenriched library, clone: 4732426A12 product: keratin complex K1C15_MOUSEKeratin, type I cytoskeletal 15 (Cytokeratin-15) (CK-15) (Keratin-15)(K15). - Mus musculus (Mouse). Q3TWV0_MOUSE Osteoclast-like cell cDNA,RIKEN full-length enriched library, clone: I420023H06 product: vimentin,full insert sequence. - Mus musculus (Mouse). Q9JKB4_MOUSE Epidermalkeratin 10 (Fragment). - Mus musculus (Mouse). Q8VCW2_MOUSE RIKEN cDNA4631426H08. - Mus musculus (Mouse). JQ0028 cytokeratin 19 - mouse KRMSE1keratin, 59K type I cytoskeletal - mouse JC4030 DnaJ-like protein MTJ1 -mouse E2AK4_MOUSE Eukaryotic translation initiation factor 2-alphakinase 4 (EC 2.7.11.1) (GCN2-like protein) (mGCN2). - Mus musculus(Mouse). BAB27580 AK011380 NID: - Mus musculus

1. A method of quantifying the amount of ionizing radiation to which anindividual has been exposed comprising determining the presence of oneor more radiation associated markers.
 2. A method according to claim 1wherein the sample is selected from the group consisting of a buccalswab, saliva, plasma, serum, urine and blood.
 3. A method according toclaim 1 wherein determining the presence of one or more radiationassociated markers in a sample is performed by Matrix assisted laserdesorption ionization (MALDI) mass spectrometry.
 4. A method accordingto claim 1 wherein determining the presence of one or more radiationassociated markers in a sample is performed by (a) contacting the samplewith an antibody which specifically binds to a radiation associatedmarker permitting formation of a complex between the antibody and theradiation associated marker; and (b) measuring the amount of complexesformed, thereby determining the amount of the radiation associatedmarker in the sample.
 5. A method according to claim 4 furthercomprising (c) comparing the amount of radiation associated marker inthe sample determined in step (b) with either (i) the amount determinedfor temporally matched, normal samples or (ii) the amount determined forsamples obtained from individuals who have not been exposed to elevatedlevels of ionizing radiation.
 6. (canceled)
 7. (canceled)
 8. (canceled)9. (canceled)
 10. (canceled)
 11. (canceled)
 12. (canceled) 13.(canceled)
 14. (canceled)